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Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

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Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

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Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
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Применить Всего найдено 5061. Отображено 100.
08-03-2012 дата публикации

Inhibition of map4k4 through rnai

Номер: US20120059046A1
Автор: Anastasia Khvorova, Joanne Kamens, Tod M Woolf, William Salomon
Принадлежит: RXi Pharmaceuticals Corp

RNAi constructs directed to MAP4K4 that demonstrate unexpectedly high gene silencing activities, and uses thereof are disclosed. The blunt-ended constructs have a double-stranded region of 19-49 nucleotides. The constructs have selective minimal modifications to confer an optimal balance of biological activity, toxicity, stability, and target gene specificity. For example, the strands may be modified (e.g., one or both ends of the sense strand is modified by 2′-O-methyl groups), such that the construct is not cleaved by Dicer or other RNAse III, the antisense strand may also be modified by a 2′-O-methyl group at the penultimate 5′-end nucleotide to greatly reduce off-target silencing.

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Номер записи: 1
24-05-2012 дата публикации

Methods and compositions for protein labeling using lipoic acid ligases

Номер: US20120129159A1
Автор: Alice Y. Ting, Daniel Shao-Chen Liu, David Baker, Florian Richter, Lucas Nivon
Принадлежит: Individual

The present disclosure provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ lipoic acid ligase or mutants thereof, and lipoic acid analogs (e.g., lipoic acid analogs comprising a resorufin moiety) recognized by lipoic acid ligase and lipoic acid ligase mutants. Also provided herein is a method of imaging protein-protein interaction via a reaction mediated by lipoic acid ligase.

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Номер записи: 2
06-12-2012 дата публикации

Thermophilic thermoanaerobacter italicus subsp. marato having high alcohol productivity

Номер: US20120309065A1
Автор: Marie Just Mikkelsen, Rasmus Lund ANDERSEN, Thomas Kvist
Принадлежит: BIOGASOL IPR APS

Strict anaerobic thermophilic bacterium belonging to the group of Thermoanaerobacter italicus subsp. marato subsp. nov. and mutants and derivatives thereof. The bacterium is particularly suitable for the production of fermentation products such as ethanol, lactic acid, acetic acid and hydrogen from lignocellulosic biomass.

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Номер записи: 3
10-01-2013 дата публикации

Compositions And Methods For Inhibiting Expression Of GSK-3 Genes

Номер: US20130012572A1
Автор: Dinah Wen-Yee Sah, Gregory Hinkle
Принадлежит: Individual

The invention relates to a double-stranded ribonucleic acid (dsRNA) targeting Glycogen Synthase Kinase-3 (GSK-3), and methods of using the dsRNA to inhibit expression of GSK-3.

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Номер записи: 4
07-02-2013 дата публикации

Modified melk peptides and vaccines containing the same

Номер: US20130034574A1
Автор: Ryuji Ohsawa, Sachiko Yoshimura, Takuya Tsunoda, Tomohisa Watanabe, Yusuke Nakamura
Принадлежит: ONCOTHERAPY SCIENCE INC

Isolated peptides composed of the amino acid sequence of the modified MELK epitope peptide or immunologically active fragments thereof that bind to HLA antigens and have higher cytotoxic T lymphocyte (CTL) inducibility than that of the wild type MELK epitope peptide and thus are suitable for use in the context of cancer immunotherapy or endometriosis immunotherapy, more particularly cancer or endometriosis vaccines are described herein. The present invention further provides peptides that include one, two, or several amino acid insertions, substitutions or additions to the aforementioned peptides or fragments, but yet retain the requisite cytotoxic T cell inducibility. Further provided are nucleic acids encoding any of these aforementioned peptides as well as pharmaceutical substances and compositions including any of the aforementioned peptides or nucleic acids. The peptides, nucleic acids, pharmaceutical substances and compositions of this invention find particular utility in the treatment of cancers, tumors, and endometriosis.

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Номер записи: 5
14-03-2013 дата публикации

Nlk as a marker for diagnosis of liver cancer and as a therapeutic agent thereof

Номер: US20130065945A1
Автор: Suk Woo Nam
Принадлежит: Industry Academic Cooperation Foundation of Catholic University of Korea

A novel marker for diagnosis of liver cancer and use thereof are provided. To be specific, a marker for diagnosis of liver cancer using over-expression of NLK (neuro-like kinase) in liver cancer cell is provided, along with a composition for diagnosis of liver cancer, a kit, a microarray, and a method for diagnosing liver cancer using the marker. Additionally, a method for screening a substance to prevent or treat liver cancer by decreasing expression of the marker gene or protein, and a composition for preventing or treating liver cancer including such substance are provided. Accordingly, the NLK gene can be efficiently used as a target for diagnosis and treatment of liver cancer.

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Номер записи: 6
04-04-2013 дата публикации

Transformed cells that ferment-pentose sugars and methods of their use

Номер: US20130084617A1
Автор: Christiaan Van Der Drift, Harry Ramancedj Harhangi, Hubertus Johannes Marie Op Den Camp, Jacobus Thomas Pronk
Принадлежит: C5 Yeast Co BV

The present invention relates to host cells transformed with a nucleic acid sequence encoding a eukaryotic xylose isomerase obtainable from an anaerobic fungus. When expressed, the sequence encoding the xylose isomerase confers to the host cell the ability to convert xylose to xylulose which may be further metabolized by the host cell. Thus, the host cell is capable of growth on xylose as carbon source. The host cell preferably is a eukaryotic microorganism such as a yeast or a filamentous fungus. The invention further relates to processes for the production of fermentation products such as ethanol, in which a host cell of the invention uses xylose for growth and for the production of the fermentation product. The invention further relates to nucleic acid sequences encoding eukaryotic xylose isomerases and xylulose kinases as obtainable from anaerobic fungi.

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Номер записи: 7
14-11-2013 дата публикации

Glycogen synthase kinase-3 inhibitors

Номер: US20130303441A1
Автор: Hagit Eldar-Finkelman, Miriam Eisenstein
Принадлежит: Ramot at Tel Aviv University Ltd, Yeda Research and Development Co Ltd

Novel peptide inhibitors of GSK-3, compositions containing same and uses thereof are disclosed. The novel peptide inhibitors are substrate-competitive inhibitors and have an amino acid sequence designed so as to bind to a defined binding site subunit in GSK-3.

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Номер записи: 8
02-01-2014 дата публикации

COMPOSITIONS AND METHODS FOR INHIBITING THE ACTIVITY OF P110a MUTANT PROTEINS

Номер: US20140005119A1
Автор: Chao Wang, Weiping Zheng, Yujun Hao, Zhengne Wang
Принадлежит: CASE WESTERN RESERVE UNIVERSITY

A method of inhibiting the activity, signaling, and/or function of a p110α mutant protein in a cancer cell expressing the p110α mutant protein includes administering to the cancer cell an amount of a therapeutic agent effective to inhibit binding of the p110α mutant protein to IRS1 in the cell.

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Номер записи: 9
03-01-2019 дата публикации

METHODS FOR TREATING CARDIAC CONDITIONS

Номер: US20190000890A1
Автор: Burrows, III Hubbard Frank, Koob Thomas J.
Принадлежит: MiMedx Group, Inc.

Described herein are compositions and methods of treating a cardiac condition using modified placental tissue or an extract of a placental tissue, capable of recruiting stem cells or promoting healing in vivo and in vitro. 1. A composition n comprising placental tissue configured for non-obstructive placement to an area approximate to a damaged cardiac tissue in an amount sufficient to reduce damage and induce healing.2. The composition of claim 1 , wherein the modified placental tissue retains an effective amount of stem cell recruiting factors claim 1 , growth factors claim 1 , and/or angiogenesis inducing factors.3. The composition of claim 1 , wherein the modified placental tissue comprises one or more of PDGF-AA claim 1 , PDGF-BB claim 1 , TGFa claim 1 , TGFB claim 1 , bFGF claim 1 , EGF claim 1 , VEGF claim 1 , IL-10 claim 1 , IL-4 claim 1 , P1GF claim 1 , TIMP-1 claim 1 , TIMP-2 claim 1 , and TIMP-4.4. The composition of any one of - claim 1 , which is a patch.5. The composition of any one of - claim 1 , wherein the modified placental tissue is micronized.6. The composition of any one of - claim 1 , which is in an injectable form.7. A method for treating injured or diseased cardiac tissue claim 1 , which method comprises placing an effective amount of a modified placenta tissue or an extract of a placental tissue at or adjacent to the injured or diseased cardiac tissue without obstructing the function thereof claim 1 , wherein the modified placenta tissue or extract is placed under conditions that promote treatment of the disease or healing of the injured or diseased cardiac tissue.8. The method of claim 7 , wherein the injured or diseased cardiac tissue is a result of ischemia claim 7 , acute myocardial infarction claim 7 , myocardial infarction claim 7 , cardiomyopathy claim 7 , unstable angina claim 7 , refractory angina claim 7 , heart attack claim 7 , heart failure claim 7 , corpulmonale claim 7 , vein graft diseases claim 7 , coronary heart diseases ...

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Номер записи: 10
06-01-2022 дата публикации

POLYPEPTIDE WITH FUNCTION OF TARGETING RECOGNITION OF IMMUNE CELLS AND APPLICATION THEREOF

Номер: US20220002686A1
Автор: "WEI Yuanan", LIU Xueshi
Принадлежит:

The present disclosure relates to a polypeptide recognizing immune cells, the polypeptide includes the following amino acid sequences: (a) an amino acid sequence containing C-terminal fragment sequence EQPDPGAVAAAAILRAILE of human Triokinase/FMN cyclase and its homologous sequence; or (b) an amino acid sequence that is substantially identical to the amino acid sequence described in (a), the substantially identical means 70% or more sequence identity to the amino acid sequence described in (a). The present invention also relates to a nucleic acid sequence encoding the polypeptide; a polypeptide probe used for targeting recognition of immune cells and containing the polypeptide described above and a reporter; a kit containing the probe described above; and, related applications of the polypeptide or probe described above. 115.-. (canceled)16. A polypeptide for targeting recognition of immune cells , comprising:(a) an amino acid sequence containing a C-terminal fragment sequence EQPDPGAVAAAAILRAILE (SEQ ID NO.:1) of human Triokinase/FMN cyclase; or(b) an amino acid sequence that is substantially identical to the amino acid sequence described in (a), wherein the substantially identical means that an amino acid sequence has more than 80% sequence identity to the amino acid sequence described in (a),wherein the amino acid sequences of (a) and (b) comprise 80 or less amino acid residues.17. The polypeptide according to claim 16 , wherein the amino acid sequences of (a) and (b) comprise 60 or less amino acid residues.18. The polypeptide according to claim 17 , wherein the amino acid sequences of (a) and (b) comprise 45 or less amino acid residues.19. The polypeptide according to claim 16 , wherein the amino acid sequence of (a) is EQPDPGAVAAAAILRAILE (SEQ ID NO.:1) claim 16 , LEQPDPGAVAAAAILRAILE (SEQ ID NO.:2) claim 16 , EQPDPGAVAAAAILRAILEVLQS (SEQ ID NO.:3) claim 16 , KNMEAGAGRASYISSARLEQPDPGAVAAAAILRAIL (SEQ ID NO.:4) claim 16 , or ...

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Номер записи: 11
04-01-2018 дата публикации

Single-Vector Gene Construct Comprising Insulin and Glucokinase Genes

Номер: US20180000967A1
Автор: BOSCH TUBERT Fatima
Принадлежит:

The invention relates to a viral expression construct and related viral vector and composition and to their use wherein said construct and vector comprise elements a) and b): a) a nucleotide sequence encoding an insulin operably linked to a first promoter, b) a nucleotide sequence encoding a glucokinase operably linked to a second promoter and said viral expression construct and related viral vector comprise at least one of elements c), d) and e): c) the first and the second promoters are positioned in reverse orientation within the expression construct, d) the first and the second promoters are positioned in reverse orientation within the expression construct and are located adjacent to each other and e) the first promoter is a CMV promoter, preferably a mini CMV promoter. 1. A viral expression construct comprising the elements a) and b):a) a nucleotide sequence encoding an insulin operably linked to a first promoter,b) a nucleotide sequence encoding a glucokinase operably linked to a second promoter and said viral expression construct comprising at least one of elements c), d) and e):c) the first and the second promoters are positioned in reverse orientation within the expression construct,d) the first and the second promoters are positioned in reverse orientation within the expression construct and are located adjacent to each other ande) the first promoter is a CMV promoter.2. A viral expression construct according to claim 1 , wherein said construct comprises elements a) claim 1 , b) and d) or wherein said construct comprises elements a) claim 1 , b) and e) wherein the first promoter is a mini CMV promoter.3. A viral expression construct according to claim 1 , wherein the first promoter is a CMV promoter claim 1 , and/or wherein the second promoter is a RSV promoter.4. A viral expression construct according to claim 1 , wherein an additional sequence is present which is selected from the group consisting of: ITRs claim 1 , SV40 polyadenylation signal claim 1 , ...

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Номер записи: 12
02-01-2020 дата публикации

NOVEL PEPTIDES AND COMBINATION OF PEPTIDES FOR USE IN IMMUNOTHERAPY AGAINST VARIOUS CANCERS

Номер: US20200000902A1
Автор: Fritsche Jens, MAHR Andrea, SCHOOR Oliver, SINGH Harpreet, SONG Colette, WEINSCHENK Toni
Принадлежит:

The present invention relates to peptides, proteins, nucleic acids and cells for use in immunotherapeutic methods. In particular, the present invention relates to the immunotherapy of cancer. The present invention furthermore relates to tumor-associated T-cell peptide epitopes, alone or in combination with other tumor-associated peptides that can for example serve as active pharmaceutical ingredients of vaccine compositions that stimulate anti-tumor immune responses, or to stimulate T cells ex vivo and transfer into patients. Peptides bound to molecules of the major histocompatibility complex (MHC), or peptides as such, can also be targets of antibodies, soluble T-cell receptors, and other binding molecules. 1. A method of treating a patient who has cancer , comprising administering to the patient an effective amount of an antibody specifically binding to an MHC class I or II molecule complexed with an antigen consisting of the amino acid sequence of VQLDSIEDLEV (SEQ ID NO: 32) ,wherein said cancer is selected from the group consisting of glioblastoma, breast cancer, colorectal cancer, renal cell carcinoma, chronic lymphocytic leukemia, hepatocellular carcinoma, non-small cell lung cancer, small cell lung cancer, Non-Hodgkin lymphoma, acute myeloid leukemia, ovarian cancer, pancreatic cancer, prostate cancer, esophageal cancer, gallbladder cancer and cholangiocarcinoma, melanoma, gastric cancer, urinary bladder cancer, head- and neck squamous cell carcinoma, and uterine cancer.2. The method of claim 1 , wherein the antibody is a polyclonal antibody claim 1 , a monoclonal antibody claim 1 , a bi-specific antibody claim 1 , or a chimeric antibody.3. The method of claim 1 , wherein the antibody binds to the HLA-restricted antigen with a binding affinity of below 20 nanomolar.4. The method of claim 1 , wherein the antibody binds to the MHC class I molecule complexed with the HLA-restricted antigen.5. The method of claim 1 , wherein the antibody is humanized.6. The ...

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Номер записи: 13
06-01-2022 дата публикации

Construction and Application of Engineered Strain of Escherichia Coli for Producing Malic Acid by Fixing CO2

Номер: US20220002766A1
Автор: Danlei MA, Guipeng HU, Jia Liu, Liming Liu, Qiuling Luo, Xiulai CHEN
Принадлежит: JIANGNAN UNIVERSITY

The disclosure discloses construction and application of an engineered strain of E. coli for producing malic acid by fixing CO2, and belongs to the field of fermentation. The engineered strain is obtained by performing genetic engineering transformation on Escherichia coli MG1655; the genetic engineering transformation includes knocking out a fumarate reductase gene, a fumarase gene, a lactate dehydrogenase gene and an alcohol dehydrogenase gene and freely overexpressing a formate dehydrogenase, an acetyl coenzyme A synthetase, an acylated acetaldehyde dehydrogenase, a formaldehyde lyase, a dihydroxyacetone kinase, a malic enzyme and a phosphite oxidoreductase to obtain a strain GH0407. The strain is used for producing malic acid by fermentation, anaerobic fermentation is performed for 72 hours with CO2 and glucose as a co-substrate, the production of malic acid reaches 39 g/L, the yield is 1.53 mol/mol, and accumulation of malic acid in the original strain is not achieved.

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Номер записи: 14
07-01-2016 дата публикации

GENES FOR IMPROVING NUTRIENT UPTAKE AND ABIOTIC STRESS TOLERANCE IN PLANTS

Номер: US20160002648A1
Автор: Guo Mei, HAYES Kevin, PETERSON-BURCH BROOKE, Simmons Carl, SIVASANKAR SOBHANA, ZOU Jijun
Принадлежит:

The present disclosure provides methods to increase crop yield utilizing transgenic genes controlling plant growth and yield. The specific genes can be used to increase tissue growth and abiotic stress tolerance. Plants, plant progeny, seeds and tissues created by these methods are also described. Polynucleotides encoding the sequences are provided for expression in a plant of interest. Expression cassettes, plants, plant cells, plant parts and seeds comprising the sequences of the disclosure are further provided. In specific embodiments, the polynucleotide is operably linked to a constitutive promoter. 1. An isolated polynucleotide selected from the group consisting of:a. a polynucleotide having at least 70% sequence identity, as determined by the GAP algorithm under default parameters, to the full length sequence of a polynucleotide selected from the group consisting of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463 ...

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Номер записи: 15
07-01-2016 дата публикации

Production of isoprene, isoprenoid, and isoprenoid precursors using an alternative lower mevalonate pathway

Номер: US20160002672A1
Автор: Dmitrii V. Vaviline, Gregory M. Whited, Guido Meurer, Jorg Mampel, Karl J. Sanford, Michael C. Miller, Walter Weyler, Zachary Q. Beck
Принадлежит: DANISCO US INC, Goodyear Tire and Rubber Co

The invention provides for compositions and methods for the production of isoprene, isoprenoid precursor, and/or isoprenoids in cells via the expression (e.g., heterologous expression) of phosphomevalonate decarboxylases and/or isopentenyl kinases.

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Номер записи: 16
07-01-2016 дата публикации

COMPOSITIONS AND METHODS FOR INCREASED ETHANOL PRODUCTION FROM BIOMASS

Номер: US20160002676A1
Автор: ARISTIDOU Aristos, JESSEN Holly J., Liao Hans, LUNDORFF Joshua, NEGRETE-RAYMOND Ana, SUOMINEN Pirkko, YI Jian
Принадлежит:

The present application discloses the identification of the novel xylose transporter genes KHT105 and RAG4, as well as the identification of a novel set of pentose phosphate pathway genes The present application further discloses a series of genetically modified yeast cells comprising various combinations of arabinose fermentation pathways, xylose fermentation pathways, pentose phosphate pathways, and/or xylose transporter genes, and methods of culturing these cells to produce ethanol in fermentation media containing xylose. 116-. (canceled)17. A genetically modified yeast cell comprising an active arabinose fermentation pathway , wherein said cell comprises one or more exogenous arabinose fermentation pathway genes selected from the group consisting of AI , RK , and RE genes , wherein the selected exogenous arabinose fermentation pathway gene encodes a polypeptide comprising an amino acid sequence with at least 80% sequence identity to an amino acid sequence selected from the group consisting of the amino acid sequences set forth in SEQ ID Nos: 6 , 8 , 10 , 12 , 14 , 16 , 18 , and 20.18. (canceled)19. The genetically modified yeast cell of further comprising an active xylose fermentation pathway claim 17 , wherein said cell comprises one or more exogenous xylose fermentation pathway genes selected from the group consisting of XR claim 17 , XDH claim 17 , and XK genes.20. The genetically modified yeast cell of further comprising an active xylose fermentation pathway claim 17 , wherein said cell comprises one or more exogenous xylose fermentation pathway genes selected from the group consisting of XI and XK genes.21. The genetically modified yeast cell of further comprising an active non-oxidative pentose phosphate pathway claim 17 , wherein said cell comprises one or more exogenous non-oxidative pentose phosphate pathway genes selected from the group consisting of TKL and TAL genes.2224-. (canceled)25. The genetically modified yeast cell of claim 17 , wherein the AI ...

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Номер записи: 17
05-01-2017 дата публикации

YEASTS MODIFIED TO USE CARBON DIOXIDE

Номер: US20170002368A1
Автор: Bideaux Carine, Bonnot Florence, Boutonnet Christel, Gorret Nathalie, Guillouet Stephane, Lesage Julie, Marc Jillian, Paques Frédéric, Pompon Denis
Принадлежит:

The invention relates to yeast cells modified to express a functional type I RuBisCO enzyme, and a class II phosphoribulokinase. The expression of these enzymes recreates a Calvin cycle in said yeasts in order to enable the yeasts to use carbon dioxide. 1. Transformed yeast cell , characterized in that it contains:a) an expression cassette containing a sequence encoding the RbcL subunit of a bacterial form I RuBisCO enzyme, under the transcriptional control of a suitable promoter;b) an expression cassette containing a sequence encoding the RbcS subunit of said RuBisCO enzyme, under the transcriptional control of a suitable promoter;c) an expression cassette containing a sequence encoding the specific chaperone RbcX of said RuBisCO enzyme, under the transcriptional control of a suitable promoter;d) an expression cassette containing a sequence encoding a general bacterial chaperone GroES, under the transcriptional control of a suitable promoter;e) an expression cassette containing a sequence encoding a general bacterial chaperone GroEL, under the transcriptional control of a suitable promoter.2. Cell according to claim 1 , characterized in that the chaperones RbcX claim 1 , GroES and GroEL come from at least two different organisms.3. Cell according to claim 1 , characterized in that the chaperone RbcX is a cyanobacterial chaperone.4. Cell according to claim 1 , characterized in that at least one of the general chaperones GroES and GroEL comes neither from a cyanobacterium nor from another bacterium expressing a RuBisCO complex.5. Cell according to claim 1 , characterized in that the three expression cassettes mentioned in points c) claim 1 , d) and e) of form a continuous block of genetic information.6. Cell according to claim 1 , characterized in that the expression cassettes mentioned in points c) claim 1 , d) and e) of are carried by a single episomal genetic element.7Saccharomyces cerevisiae.. Yeast cell according to claim 1 , characterized in that said yeast ...

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Номер записи: 18
05-01-2017 дата публикации

MICROORGANISMS HAVING PUTRESCINE PRODUCTIVITY AND PROCESS FOR PRODUCING PUTRESCINE USING THE SAME

Номер: US20170002386A1
Автор: JUNG Hee Kyoung, KIM Chang Gyeom, LEE Baek Seok, Lee Kyoung Min, Lee Sun Young, LI Hong Xian, Park Su Jin, Um Hye Won, YANG Young Lyeol, YOON Jong Hyun
Принадлежит: CJ CHEILJEDANG CORPORATION

The present invention relates to a recombinant microorganism capable of producing putrescine, in which the microorganism is modified to have enhanced NCgl2522 activity, thereby producing putrescine in a high yield, and a method for producing putrescine using the microorganism. 1. A microorganism having putrescine productivity , which is modified to have enhanced activity of a protein having an amino acid sequence represented by SEQ ID NO: 21 or 23.2. The microorganism having putrescine productivity according to claim 1 , wherein the microorganism is further modified to have weakened activities of ornithine carbamoyltransferase (ArgF) and a protein (NCgl1221) involved in glutamate export claim 1 , compared to the endogenous activities claim 1 , and to have enhanced ornithine decarboxylase (ODC) activity.3. The microorganism having putrescine productivity according to claim 2 , wherein the ornithine carbamoyltransferase (ArgF) has an amino acid sequence represented by SEQ ID NO: 29 claim 2 , the protein (NCgl1221) involved in glutamate export has an amino acid sequence represented by SEQ ID NO: 30 claim 2 , and the ornithine decarboxylase (ODC) has an amino acid sequence represented by SEQ ID NO: 33.4. The microorganism having putrescine productivity according to claim 1 , wherein the microorganism is further modified to have enhanced activities of acetyl-gamma-glutamyl-phosphate reductase (ArgC) claim 1 , acetylglutamate synthase or ornithine acetyltransferase (ArgJ) claim 1 , acetylglutamate kinase (ArgB) claim 1 , and acetylornithine aminotransferase (ArgD) claim 1 , compared to the endogenous activities.5. The microorganism having putrescine productivity according to claim 4 , wherein the acetyl-gamma-glutamyl-phosphate reductase (ArgC) claim 4 , acetylglutamate synthase or ornithine acetyltransferase (ArgJ) claim 4 , acetyl glutamate kinase (ArgB) claim 4 , and acetylornithine aminotransferase (ArgD) have amino acid sequences represented by SEQ ID NOs: 25 claim 4 ...

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Номер записи: 19
05-01-2017 дата публикации

MOLECULE ASSOCIATED WITH ONSET OF GOUT, AND METHOD AND KIT FOR EVALUATING DIATHESIS OF URIC ACID-RELATED DISEASES AND INFLAMMATION-RELATED DISEASES, AND INSPECTION OBJECT AND DRUG

Номер: US20170002413A1
Автор: Matsuo Hirotaka, Shinomiya Nariyoshi, Takada Tappei
Принадлежит:

To specify a molecule associated with the onset of gout so as to provide a method for evaluating a diathesis of uric acid-related diseases and a diathesis of inflammation-related diseases, an evaluation kit for carrying out the method, an inspection object, and a drug, on the basis of the molecule specified above, for contributing to the early treatment and prevention of the uric acid-related diseases and inflammation-related diseases. The molecule includes any one protein and cDNA of CNIH2-PACS1, ALDH2, MYL2-CUX2, GCKR, MAP3K11, NPT4, ABCG2, HIST1H2BF/HIST1H4E, HIST1H2BE/HIST1H4D and FAM35A, or proteins of combination thereof with GLUT9, NPT1, URAT1, or NXRN2, and is capable of selectively inducing gout. A molecule includes protein and cDNA of an ABCG2 variant and is capable of selectively and ATP-dependently decreasing urate excretion. 1. A molecule associated with onset of gout , comprising any one protein or cDNA of CNIH2-PACS1 , ALDH2 , MYL2-CUX2 , GCKR , MAP3K11 , NPT4 , ABCG2 , HIST1H2BF/HIST1H4E , HIST1H2BE/HIST1H4D , and FAM35A , or a combination thereof with any one protein or cDNA of GLUT9 , NPT1 , URAT1 , and NXRN2 , and being capable of relating to the onset of gout; or comprising protein or cDNA of an ABCG2 variant , and being capable of selectively and ATP-dependently decreasing excretion of urate.2. A method for evaluating a uric acid-related disease diathesis and an inflammation-related disease diathesis , the method comprising:evaluating whether or not a subject has a diathesis capable of inducing urate regulation failure, or a state or a uric acid-related disease attributable to the failure, and the evaluating comprising:a step of detecting a gene polymorphism of a gene encoding at least any one protein or cDNA of CNIH2-PACS1, ALDH2, MYL2-CUX2, GCKR, MAP3K11, NPT4, ABCG2, HIST1H2BF/HIST1H4E, HIST1H2BE/HIST1H4D, and FAM35A, or a combination thereof with gene polymorphisms of GLUT9, NPT1, URAT1, and NXRN2, using a test sample containing human genes ...

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Номер записи: 20
02-01-2020 дата публикации

MICROORGANISMS FOR PRODUCING PUTRESCINE OR ORNITHINE AND PROCESS FOR PRODUCING PUTRESCINE OR ORNITHINE USING THEM

Номер: US20200002686A1
Автор: JUNG Hee Kyoung, LEE Baek Seok, LEE Hyo Hyoung, Lee Kyoung Min, LI Hong Xian, Park Su Jin, Um Hye Won, YANG Young Lyeol
Принадлежит:

Disclosed is a modified microorganism producing putrescine or ornithine, and a method for producing putrescine or ornithine using the same. 115-. (canceled)16. A method for producing putrescine or ornithine , comprising:{'i': Corynebacterium', 'E. coli', 'E. coli, '(i) culturing a modified microorganism of the genus producing putrescine or ornithine in a medium, wherein activities of N-acetylglutamate synthase from and acetylornithine deacetylase from are introduced into the microorganism; and'}(ii) recovering putrescine or ornithine from the cultured microorganism or the medium.17CorynebacteriumCorynebacterium glutamicum.. The method according to claim 16 , wherein the microorganism of the genus is18E. coliE. coli. The method according to claim 16 , wherein the N-acetylglutamate synthase from consists of an amino acid sequence of SEQ ID NO: 1 claim 16 , and/or the acetylornithine deacetylase from consists of an amino acid sequence of SEQ ID NO: 3.19. The method according to claim 16 , wherein (a) an activity of phosphotransacetylase and acetate kinase operon (pta-ackA operon); (b) an activity of at least one selected from the group consisting of acetyl gamma glutamyl phosphate reductase (ArgC) claim 16 , acetylglutamate synthase or ornithine acetyltransferase (ArgJ) claim 16 , acetylglutamate kinase (ArgB) claim 16 , and acetyl ornithine aminotransferase (ArgD); and/or (c) an activity of putrescine exporter is further enhanced compared to its endogenous activity.20E. coli. The method according to claim 16 , wherein an activity of acetyl-CoA synthetase (acs) from claim 16 , and/or an activity of ornithine decarboxylase (ODC) is further introduced.21. The method according to claim 16 , wherein (a) an activity of i) ornithine carbamoyltransferase (ArgF) claim 16 , ii) glutamate exporter claim 16 , or iii) ornithine carbamoyltransferase and glutamate exporter and/or (b) an activity of acetyltransferase is further weakened compared to its endogenous activity. This ...

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Номер записи: 21
02-01-2020 дата публикации

Variants of a DNA Polymerase of the Polx Family

Номер: US20200002690A1
Автор: Delarue Marc, Ybert Thomas
Принадлежит:

The invention relates to variants of a DNA polymerase of the polX family capable of synthesizing a nucleic acid molecule without a template strand, or of a functional fragment of such a polymerase, comprising at least one mutation of a residue in at least one specific position, and to uses of said variants, in particular for the synthesis of nucleic acid molecules comprising 3′-OH modified nucleotides. 124-. (canceled)25. A DNA polymerase of the polX family capable of synthesizing a nucleic acid molecule without a template strand , said DNA polymerase comprising at least one mutation of a residue in at least one position selected from the group consisting of E457 , T331 , G332 , G333 , F334 , R336 , K338 , H342 , D343 , V344 , D345 , F346 , A397 , D399 , D434 , V436 , A446 , L447 , L448 , G449 , W450 , G452 , R454 , Q455 , F456 , R458 , R461 , N474 , E491 , D501 , Y502 , 1503 , P505 , R508 , N509 and A510 , the positions indicated being determined by alignment with SEQ ID No. 1.26. The DNA polymerase of the polX family according to claim 25 , said DNA polymerase being capable of synthesizing a DNA strand and/or an RNA strand.27. The DNA polymerase of the polX family according to claim 25 , said DNA polymerase being a variant of Pol IV claim 25 , Pol μ claim 25 , or of the terminal deoxyribonucleotidyl transferase (TdT).28. The DNA polymerase of the polX family according to claim 25 , in which at least one mutation consists of a substitution claim 25 , a deletion or an addition of one or more amino acid residues.29. The DNA polymerase of the polX family according to claim 25 , said DNA polymerase comprising at least one mutation of a residue in at least one position selected from the group consisting of T331 claim 25 , G332 claim 25 , G333 claim 25 , F334 claim 25 , R336 claim 25 , D343 claim 25 , L447 claim 25 , L448 claim 25 , G449 claim 25 , W450 claim 25 , G452 claim 25 , R454 claim 25 , Q455 claim 25 , E457 claim 25 , R461 and R508.30. The DNA polymerase of the ...

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Номер записи: 22
07-01-2021 дата публикации

COMPOSITIONS AND METHODS INVOLVING ENGINEERED P27

Номер: US20210002619A1
Автор: GUILEY Keelan, RUBIN Seth
Принадлежит: The Regents of the University of California

The disclosure provides polypeptides comprising an engineered p27, or a fragment thereof. Such polypeptides may be used to form trimeric protein complexes with a cyclin-dependent kinase 4 (Cdk4) (or a variant thereof) or Cdk6 (or a variant thereof), and a cyclin D (CycD) or a variant thereof. 1. A polypeptide comprising an engineered p27 , or a fragment thereof , wherein the engineered p27 has at least one amino acid substitution at a position selected from the group consisting of Y74 , Y88 , and Y89 , wherein the engineered p27 forms a trimeric protein complex with (i) a cyclin-dependent kinase 4 (Cdk4) or a variant thereof , or a Cdk6 or a variant thereof , and (ii) a cyclin D (CycD) or a variant thereof , and wherein the amino acid positions are determined with reference to the sequence of SEQ ID NO: 1.2. The polypeptide of claim 1 , wherein the amino acid substitution at position Y74 is Y74E claim 1 , Y74D claim 1 , or Y74R claim 1 , the amino acid substitution at Y88 is Y88E or Y88D claim 1 , and the amino acid substitution at Y89 is Y89E or Y89D.34-. (canceled)5. The polypeptide of claim 1 , wherein the engineered p27 comprises a sequence having at least 90% sequence identity to the sequence of SEQ ID NO: 1.716-. (canceled)17. The polypeptide of claim 6 , wherein the engineered p27 comprises a sequence having at least 90% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 6 claim 6 , 4 claim 6 , 12 claim 6 , 10 claim 6 , 15 claim 6 , 13 claim 6 , 21 claim 6 , 19 claim 6 , 27 claim 6 , 25 claim 6 , 30 claim 6 , 28 claim 6 , 33 claim 6 , 31 claim 6 , 36 claim 6 , and 34.1824-. (canceled)25. A trimeric protein complex comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(i) the polypeptide of , or a phosphorylated, wild-type p27 or a fragment thereof;'}(ii) a Cdk4 or a variant thereof, or a Cdk6 or a variant thereof; and(iii) a CycD or a variant thereof,wherein the Cdk4 or the variant thereof or the Cdk6 or the variant ...

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Номер записи: 23
07-01-2021 дата публикации

MODIFIED AGPASE LARGE SUBUNIT SEQUENCES AND METHODS FOR DETECTION OF PRECISE GENOME EDITS

Номер: US20210002658A1
Автор: Begemann Matthew, January Emma
Принадлежит: BENSON HILL, INC.

Compositions and methods for improving plant growth are provided herein. Polynucleotides encoding modified AGPase large subunit proteins, polypeptides encompassing modified AGPase large subunit proteins, methods of producing modified polynucleotides encoding modified AGPase large subunit proteins, and expression constructs for expressing genes of interest whose expression may improve agronomic properties including but not limited to crop yield, biotic and abiotic stress tolerance, and early vigor, plants comprising the polynucleotides, polypeptides, and expression constructs, and methods of producing plants comprising the modified AGPase large subunit genes of the invention or encoding the AGPase large subunit proteins of the invention are also provided. Compositions and methods for the ready detection of precise base changes are also provided herein. Compositions include primer pads as part of a repair donor template, and methods for the detection of precise base changes include PCR detection using one or more primers designed to anneal with a primer pad sequence. 2. The modified AGPase large subunit protein of wherein said AGPase large subunit protein shares at least 80% identity with SEQ ID NO:6 claim 1 , 8 claim 1 , 10 claim 1 , 12 claim 1 , 14 claim 1 , or 16 claim 1 , or is encoded by a polynucleotide that shares at least 70% identity with a sequence selected from the group consisting of SEQ ID NOs:5 claim 1 , 7 claim 1 , 9 claim 1 , 11 claim 1 , 13 claim 1 , 15 claim 1 , and 43-48.4. The modified AGPase large subunit protein of wherein said AGPase large subunit protein shares at least 80% identity with a sequence selected from the group consisting of SEQ ID NOs:49-54 claim 3 , or is encoded by a polynucleotide that shares at least 70% identity with a sequence selected from the group consisting of SEQ ID NOs:55-66.5. The modified AGPase large subunit protein of wherein said modified AGPase large subunit has at least 80% sequence identity to a sequence selected ...

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Номер записи: 24
03-01-2019 дата публикации

Double-stranded oligonucleotides

Номер: US20190002880A1
Автор: Kristin Wiederholt, Tod Woolf
Принадлежит: Life Technologies Corp

Antisense sequences, including duplex RNAi compositions, which possess improved properties over those taught in the prior art are disclosed. The invention provides optimized antisense oligomer compositions and method for making and using the both in in vitro systems and therapeutically. The invention also provides methods of making and using the improved antisense oligomer compositions.

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Номер записи: 25
03-01-2019 дата публикации

BACTERIAL CELLS WITH IMPROVED TOLERANCE TO POLYAMINES

Номер: US20190002935A1
Автор: Feist Adam, Herrgard Markus, Lennen Rebecca, Mohamed Elsayed Tharwat Tolba, Nielsen Alex Toftgaard, SOMMER Morten
Принадлежит:

Provided are bacterial cells genetically modified to improve their tolerance to certain commodity chemicals, such as polyamines, and methods of preparing and using such bacterial cells for production of polyamines and other compounds. 1. A bacterial cell comprising a recombinant biosynthetic pathway for producing an aliphatic polyamine and at least one genetic modification which reduces expression of an endogenous gene selected from the group consisting of proV , proW , proX , cspC , ptsP , wbbK , yobF , nagC , nagA , rph , ybeX and mpl , or a combination of any thereof.2. The bacterial cell of claim 1 , comprising a genetic modification which reduces expression of ybeX claim 1 , proV claim 1 , cspC claim 1 , ptsP claim 1 , wbbK claim 1 , mpl or rph.3. The bacterial cell of claim 2 , comprising genetic modifications which reduce the expression ofa) proV and at least one of ptsP, cspC, mpl, and ybeX;b) proV, ptsP, and at least one of mpl and ybeX;c) proV, cspC, and at least one of mpl and ybeX;d) ybeX and at least one of proV, ptsP, cspC, and mpl;e) proV, ptsP, ybeX, and mpl; orf) proV, cspC, ybeX, and mpl.4. The bacterial cell of claim 1 , wherein the genetic modification comprises a knock-down or knock-out of the endogenous gene.5. The bacterial cell of claim 1 , further comprising a mutation in at least one of YgaC claim 1 , RpsG claim 1 , MreB claim 1 , NusA claim 1 , SspA claim 1 , MrdB claim 1 , RpoD claim 1 , RpoC claim 1 , RpoB claim 1 , MurA claim 1 , RpsA claim 1 , SpoT claim 1 , argG claim 1 , rph or the pyrE/rph intergenic region.6. A bacterial cell comprising at least one mutation selected from YgaC-R43L claim 1 , RpsG-L157* claim 1 , MreB-A298V claim 1 , MreB-N34K claim 1 , MreB-E212A claim 1 , MreB-I24M claim 1 , MreB-H93N claim 1 , NusA-L152R claim 1 , NusA-M204R claim 1 , SspA-F83C claim 1 , SspA-V91F claim 1 , MrdB-E254K claim 1 , RpoD-E575A claim 1 , RpoC-V401G claim 1 , RpoC-V453I claim 1 , RpoC-R1140C claim 1 , RpoC-L120P claim 1 , RpoB-R637L ...

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Номер записи: 26
13-01-2022 дата публикации

GLYCOGEN-NULL METHANOTROPHS AND USES THEREOF

Номер: US20220010268A1
Автор: Hsi Megan, Luning Eric G., Rueda Paloma, Saville Renee M., Silverman Joshua A., Stegantseva Yelena, Tang Vincent
Принадлежит:

The present disclosure provides methanotrophic bacteria that are modified to produce less glycogen, and methods of using the modified methanotrophic bacteria to produce a desired product, such as protein(s) or metabolite(s). 1Methylococcus capsulatusMethylococcus capsulatus. A modified , comprising a chromosomal knock-out of an ADP-glucose pyrophosphorylase gene , a glgA2 isoform of a glycogen synthase gene , or both , wherein the modified cultured under conditions comprising a non-limiting amount of a Csubstrate produces:{'i': 'Methylococcus capsulatus', 'at least about 30% less glycogen as compared to the parent cultured under the same conditions; and/or'}{'i': 'Methylococcus capsulatus', 'at least about 5% more crude protein as compared to the parent cultured under the same conditions.'}2Methylococcus capsulatusMethylococcus capsulatusMethylococcus capsulatus. The modified of claim 1 , wherein the is Bath.3Methylococcus capsulatus. The modified of or claim 1 , wherein the culture conditions further comprise the presence of a limiting amount of a nutrient or metabolite required for growth.4Methylococcus capsulatus. The modified of claim 3 , wherein the limiting amount of the nutrient required for growth comprises a limiting amount of nitrogen claim 3 , sulfur claim 3 , phosphorous claim 3 , and/or oxygen.5Methylococcus capsulatus. The modified of claim 4 , wherein the limiting amount of the nitrogen comprises a limiting amount of nitrate claim 4 , ammonium claim 4 , and/or nitrogen gas.6Methylococcus capsulatusMethylococcus capsulatus. The modified of any of - claim 4 , wherein the production of at least about 5% more crude protein as compared to the parent further comprises a culture condition comprising from about 20% to about 80% nitrogen fixation.7Methylococcus capsulatusMethylococcus capsulatusMethylococcus capsulatus.. The modified of any of - claim 4 , wherein the modified has a lower ratio of utilized oxygen to utilized methane as compared to the ...

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Номер записи: 27
01-01-2015 дата публикации

Heat stable mutants of starch biosynthesis enzymes

Номер: US20150007363A1
Автор: L. Curtis Hannah, Thomas W. Greene
Принадлежит: University of Florida Research Foundation Inc

The subject invention pertains to novel mutant polynucleotide molecules that encode enzymes that have increased heat stability. These polynucleotides, when expressed in plants, result in increased yield in plants grown under conditions of heat stress. The polynucleotide molecules of the subject invention encode maize endosperm ADP glucose pyrophosphorylase (AGP) and soluble starch synthase (SSS) enzyme activities. Plants and plant tissue bred to contain, or transformed with, the mutant polynucleotides, and expressing the polypeptides encoded by the polynucleotides, are also contemplated by the present invention. The subject invention also concerns methods for isolating polynucleotides and polypeptides contemplated within the scope of the invention. Methods for increasing yield in plants grown under conditions of heat stress are also provided.

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Номер записи: 28
11-01-2018 дата публикации

Cleavable Lipids

Номер: US20180008543A1
Автор: DeRosa Frank, GUILD Braydon Charles, Heartlein Michael, Zhang Jerry Chi
Принадлежит:

Disclosed herein are novel compounds, pharmaceutical compositions comprising such compounds and related methods of their use. The compounds described herein are useful, e.g., as liposomal delivery vehicles to facilitate the delivery of encapsulated polynucleotides to target cells and subsequent iransfection of said target cells, and in certain embodiments are characterized as having one or more properties that afford such compounds advantages relative to other similarly classified lipids. 2. (canceled)3. The nanoparticle of claim 1 , wherein Ris imidazole.4. The nanoparticle of claim 1 , wherein{'sub': '1', 'Ris imidazole;'}andn is 1.5. The nanoparticle of claim 1 , wherein Ris guanidinium.6. The nanoparticle-of claim 1 , wherein{'sub': '1', 'Ris guanidinium;'}andn is 1.723.-. (canceled)2629.-. (canceled)30. The nanoparticle of claim 1 , further comprising one or more compounds selected from the group consisting of a cationic lipid claim 1 , a PEG-modified lipid claim 1 , a non-cationic lipid and a helper lipid.31. (canceled)32. The nanoparticle of claim 1 , wherein one or more of the polynucleotides comprises a chemical modification.33. The nanoparticle of claim 1 , wherein the one or more polynucleotides is selected from the group consisting of an antisense oligonucleotide claim 1 , siRNA claim 1 , miRNA claim 1 , snRNA claim 1 , snoRNA and combinations thereof.34. (canceled)35. The nanoparticle of claim 1 , wherein the one or more polynucleotides comprise DNA.36. The nanoparticle of claim 1 , wherein the one or more polynucleotides comprise RNA.37. (canceled)38. The nanoparticle of claim 36 , wherein the RNA encodes an enzyme.39. The nanoparticle of claim 38 , wherein the enzyme is selected from the group consisting of agalsidase alfa claim 38 , alpha-L-iduronidase claim 38 , iduronate-2-sulfatase claim 38 , N-acetylglucosamine-1-phosphate transferase claim 38 , N-acetylglucosaminidase claim 38 , alpha-glucosaminide acetyltransferase claim 38 , N- ...

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Номер записи: 29
12-01-2017 дата публикации

VECTORS AND STRAINS FOR PRODUCING MYRCENE AND METHOD OF PRODUCING MYRCENE USING THE SAME

Номер: US20170009240A1
Автор: GONG Gyeongtaek, KIM Eun Mi, KIM Yunje, Lee Sun Mi, UM Youngsoon, WOO Han Min
Принадлежит: KOREA INSTITUTE OF SCIENCE AND TECHNOLOGY

Disclosed herein are an expression vector capable of expressing myrcene, an strain transformed with the vector and having improved capability of producing myrcene and a method for producing myrcene and a method for recycling glycerol using the same. In an aspect, the transformed strain of the present disclosure can produce myrcene with high purity on a large scale using glycerol or glucose as a carbon source. Also, the strain of the present disclosure is economical and environment-friendly because it can produce high value-added myrcene using waste glycerol as a carbon source. In addition, the strongly volatile myrcene can be produced and isolated at the same time. 1Escherichia coli. A transformed strain transformed with a first vector and a second vector ,the first vector comprising, in sequence,a chloramphenicol resistance gene as a selection marker;a p15A replication origin as a replication origin;a lacUV5 promoter;a first domain comprising a gene encoding an enzyme which produces mevalonate from acetyl-CoA; anda second domain comprising a gene encoding an enzyme which produces dimethylallyl pyrophosphate (DMAPP) from mevalonate, andthe second vector comprising, in sequence,an ampicillin resistance gene as a selection marker;a ColE1 replication origin as a replication origin;a trc promoter; anda gene encoding an enzyme which is capable of producing myrcene from geranyl pyrophosphate (GPP).2Escherichia coli. The transformed strain according to claim 1 ,wherein the first vector further comprises one or more selected from a trc promoter; and a gene encoding an enzyme which is capable of producing geranyl pyrophosphate (GPP) from dimethylallyl pyrophosphate (DMAPP) and isopentenyl diphosphate (IPP),the trc promoter is located between the first domain and the second domain, andthe gene encoding an enzyme which is capable of producing geranyl pyrophosphate (GPP) from dimethylallyl pyrophosphate (DMAPP) and isopentenyl diphosphate (IPP) is located downstream of the ...

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Номер записи: 30
10-01-2019 дата публикации

Inhibition of Diacylglycerol Kinase to Augment Adoptive T cell Transfer

Номер: US20190008898A1
Автор: Gary Koretzky, Liang-Chuan WANG, Matthew RIESE, Steven M. Albelda
Принадлежит: University of Pennsylvania Penn

The present invention provides compositions and methods for inhibiting one or more diacylglycerol kinase (DGK) isoform in a cell in order to enhance the cytolytic activity of the cell. In one embodiment, the cells may be used in adoptive T cell transfer. For example, in some embodiments, the cell is modified to express a chimeric antigen receptor (CAR). Inhibition of DGK in T cells used in adoptive T cell transfer increases cytolytic activity of the T cells and thus may be used in the treatment of a variety of conditions, including cancer, infection, and immune disorders.

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Номер записи: 31
14-01-2016 дата публикации

Antibodies to argininosuccinate synthase and related methods

Номер: US20160009821A1
Автор: Bor-Wen Wu, Wei He, Yunyun GUO
Принадлежит: Tdw Group

Provided are antibodies, and antigen-binding fragments thereof, which specifically bind to argininosuccinate synthase, and related compositions, kits, and methods of use thereof, for instance, as companion diagnostics to identify suitable subjects for arginine deprivation or depletion therapies such as ADI-PEG 20 and other arginine deiminase (ADI) polypeptide-based therapies.

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Номер записи: 32
11-01-2018 дата публикации

CELLS GENETICALLY MODIFIED TO COMPRISE PANCREATIC ISLET GLUCOKINASE AND USES THEREOF

Номер: US20180010104A1
Автор: Simpson Ann Margaret, Tao Chang
Принадлежит:

The present invention relates generally to a population of cells genetically modified to produce insulin in a glucose responsive manner and uses thereof. More particularly, the present invention relates to a population of cells genetically modified to produce insulin in response to physiologically relevant levels of glucose and uses thereof. The cells of the present invention are useful in a wide variety of applications, in particular in the context of therapeutic and prophylactic regimes directed to the treatment of diabetes and/or the amelioration of symptoms associated with diabetes, based on the transplantation of the cells of the present invention into mammals requiring treatment. Also facilitated is the design of in vitro based screening systems for testing the therapeutic effectiveness and/or toxicity of potential adjunctive treatment regimes. 1. (canceled)2. An isolated mammalian hepatocyte recombinantly expressing insulin protein , pancreatic islet glucokinase protein , and GLUT2 protein , and wherein the mammalian hepatocyte produces insulin when exposed to an extracellular glucose concentration from about 3 mM to about 8 mM.3. The mammalian hepatocyte of claim 2 , wherein the insulin protein is a human insulin protein claim 2 , the pancreatic islet glucokinase protein is a human pancreatic islet glucokinase claim 2 , and the GLUT2 protein is a human GLUT2 protein.4. The mammalian hepatocyte of claim 2 , wherein the pancreatic islet glucokinase has an amino acid sequence of SEQ ID NO: 2 or at least 90% identical to SEQ ID NO: 2.5. The mammalian hepatocyte of claim 2 , wherein the mammalian hepatocyte is a human hepatocyte.6. The mammalian hepatocyte of claim 2 , wherein the mammalian hepatocyte is a Huh7 cell.7. The mammalian hepatocyte of claim 2 , wherein the hepatocytes are autologous cells claim 2 , allogenic cells claim 2 , or combination thereof.8. The mammalian hepatocyte of claim 2 , wherein the hepatocytes are encapsulated.9. An isolated mammalian ...

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Номер записи: 33
14-01-2021 дата публикации

MODIFIED TEMPLATE-INDEPENDENT ENZYMES FOR POLYDEOXYNUCLEOTIDE SYNTHESIS

Номер: US20210009969A1
Автор: Efcavitch J. William, Sammond Deanne W., Sinha Prem, Stec Boguslaw, Tubbs Julie L., Wilson Christopher
Принадлежит:

The invention includes methods for identifying polymerases, such as modified terminal nucleotidyl transferases (TdT), that are capable of binding nucleotides comprising removable 3′-O-blocking moieties to a nucleic acid initiator, without the use of a template. The invention further includes the identified polymerases, and methods of using the polymerases for de novo synthesis of predetermined oligonucleotide sequences. 1. A modified terminal deoxynucleotidyl transferase (TdT) comprising a mutation selected from the group consisting of E33K , E180L , E180K , M192E , M192K , M192W , W303H , L381K , L381Q , L381R , L381V , W450H , R454I , R454T , R454K , E457K , R461V , R461Q , R461V , N474R , and N474K , said modified TdT capable of adding a nucleotide analog comprising a removable blocking moiety at a 3′-Oxygen of the analog to a 3′-OH of a nucleic acid initiator in the absence of a nucleic acid template.2. The modified TdT of claim 1 , comprising a mutation E457K.3. The modified TdT of claim 1 , comprising the mutations E180K claim 1 , M192W claim 1 , L381R claim 1 , and W450H.4. The modified TdT of claim 1 , comprising the mutations L381Q and W450H.5. The modified TdT of claim 1 , comprising the mutations E180L claim 1 , M193E claim 1 , L381K claim 1 , R461Q claim 1 , and N457K.6. The modified TdT of claim 1 , comprising the mutations E180K claim 1 , L381Q claim 1 , W450H and R461V.7. The modified TdT of claim 1 , comprising the mutations L381Q and W450H.8. The modified TdT of claim 1 , comprising the mutations E180L claim 1 , M192E claim 1 , L381K claim 1 , R461Q claim 1 , and N457K.9. The modified TdT of claim 1 , comprising the mutations E180K claim 1 , M192E claim 1 , L381K claim 1 , R454T claim 1 , and N47K.10. The modified TdT of claim 1 , comprising the mutations E180K claim 1 , M192K claim 1 , L381K claim 1 , R454T claim 1 , and N457R.11. The modified TdT of claim 1 , comprising the mutations E180K claim 1 , M192K claim 1 , L381K claim 1 , R454K claim 1 , ...

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Номер записи: 34
14-01-2021 дата публикации

Variants of Terminal Deoxynucleotidyl Transferase and Uses Thereof

Номер: US20210009970A1
Автор: Elise Champion, Marc Delarue, Mikhael SOSKINE, Thomas Ybert
Принадлежит: DNA Script SAS, Institut Pasteur de Lille

The present invention relates to variants of Terminal deoxynucleotidyl Transferase (TdT), each of which (i) has an amino acid sequence similarity to SEQ ID NO: 2. 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 or 35 with corresponding amino acid substitutions, (ii) is capable of synthesizing a nucleic acid fragment without a template and (iii) is capable of incorporating a modified nucleotide into the nucleic acid fragment.

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Номер записи: 35
10-01-2019 дата публикации

Enzyme Composition for DNA End Repair, Adenylation, Phosphorylation

Номер: US20190010482A1
Автор: Berezniakovas Arturas, Lubiene Judita, LUBYS Arvydas
Принадлежит: Thermo Fisher Scientific Baltics UAB

Enzyme compositions and their method of use that provide ready-to-use master mixtures. The compositions comprise a modified thermophilic DNA polymerase lacking 5′-3′ and 3′-5′ exonuclease activity premixed with T4 DNA polymerase, Klenow fragment and T4 polynucleotide kinase and all other necessary components, including reaction buffer and nucleoside triphosphates, required to perform DNA blunting, phosphorylation, and single nucleotide extension reactions in one tube and in two steps. Among other benefits, the mixture of different enzymes, buffers and nucleoside triphosphates is stable during prolonged storage. 120-. (canceled)21. An enzyme composition comprising in a single container a buffer comprising nucleoside triphosphates and a plurality of enzymes separately capable of blunting , phosphorylating and adenylating DNA fragments , with the enzyme capable of adenylating DNA being a thermophilic modified DNA polymerase , and the composition being stable for at least one week at 4° C.22. The composition of claim 21 , where the enzymes capable of blunting and phosphorylating DNA fragments are chosen from T4 DNA polymerase claim 21 , T7 DNA polymerase claim 21 , Klenow fragment claim 21 , and T4 polynucleotide kinase.23Thermus. The composition of claim 21 , where the thermophilic modified DNA polymerase is a species thermophilic DNA polymerase fused to non-specific DNA-binding domain.24. The composition of claim 21 , where the thermophilic modified DNA polymerase lacks 5′-3′ and 3′-5′ exonuclease activity.25ThermusSulfolobus. The composition of claim 23 , where the thermophilic modified DNA polymerase is a species polymerase fused to non-specific DNA-binding domain from family.26. The composition of claim 21 , where the nucleoside triphosphates are dATP claim 21 , dCTP claim 21 , dTTP claim 21 , and dGTP claim 21 , where the concentration of dATP ranges from 0.4 to 3 mM claim 21 , the concentration of dCTP ranges from 0.2 to 0.6 mM claim 21 , the concentration of ...

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Номер записи: 36
14-01-2021 дата публикации

AMINO ACID-SENSING DIGUANYLATE CYCLASE AND METHODS OF USE

Номер: US20210010057A1
Автор: Guillemin Karen, Robinson Catherine, Sweeney Emily
Принадлежит: University of Oregon

Provided are SpdE polypeptides and variants and nucleic acids encoding the SpdE polypeptides and variants. Also provided are vectors including one or more nucleic acids encoding a SpdE polypeptide or variant and cells including a nucleic acid encoding the SpdE polypeptide or variant, as well as cells expressing a SpdE polypeptide or variant and compositions including such cells and a pharmaceutically acceptable carrier. Finally, methods of detecting presence and/or amount of one or more amino acids in a sample are provided. The methods include contacting the sample with a SpdE protein, measuring diguanylate cyclase activity of the SpdE protein; and comparing the diguanylate cyclase activity of the SpdE protein to a control. The methods can utilize isolated SpdE protein or a cell expressing a SpdE protein. 1. A method of detecting presence or amount of one or more amino acids in a sample , comprising:contacting the sample with a SpdE protein;measuring diguanylate cyclase activity of the SpdE protein; andcomparing the diguanylate cyclase activity of the SpdE protein to a control, wherein a decrease in diguanylate cyclase activity compared to the control indicates the presence or amount of one or more amino acids in the sample.2. The method of claim 1 , wherein the one or more amino acids are one or more of proline claim 1 , valine claim 1 , isoleucine claim 1 , leucine claim 1 , alanine claim 1 , methionine claim 1 , or threonine.3. The method of claim 1 , wherein measuring the diguanylate cyclase activity of the SpdE protein comprises measuring an amount of cyclic-di-GMP or pyrophosphate.4. The method of claim 1 , wherein contacting the sample with the SpdE protein comprises contacting the sample with a cell expressing the SpdE protein.5. The method of claim 4 , wherein measuring the diguanylate cyclase activity of the SpdE protein comprises measuring the motility of the cell expressing the SpdE protein and wherein an increase in the motility of the cell compared to ...

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Номер записи: 37
10-01-2019 дата публикации

BACTERIA ENGINEERED TO TREAT METABOLIC DISEASES

Номер: US20190010506A1
Автор: Falb Dean, Fisher Adam B., Isabella Vincent M., Kotula Jonathan W., Miller Paul F., MILLET Yves, Rowe Sarah Elizabeth, Tucker Alex
Принадлежит:

Genetically engineered bacteria, pharmaceutical compositions thereof, and methods of attenuating metabolic diseases are disclosed. 1bacterium. An engineered comprising a gene sequence or gene cassette for producing one or more aryl hydrocarbon receptor (AhR) agonist(s) , wherein the gene sequence or gene cassette is operably linked to a directly or indirectly inducible promoter that is not associated with the gene sequence or gene cassette in nature.2bacteriumbacterium. The engineered of claim 1 , wherein the engineered comprises gene sequence for producing indole-3-acetonitrile.3bacteriumbacterium. The engineered of claim 2 , wherein the engineered comprises gene sequence encoding cyp79B2 (tryptophan N-monooxygenase).4. The genetically engineered bacteria of or claim 2 , wherein the engineered bacterium comprises gene sequence encoding cyp71a13 (indoleacetaldoxime dehydratase).5. The genetically engineered bacteria of any of - claim 2 , wherein the engineered bacterium comprises gene sequence encoding cyp79B3 (tryptophan N-monooxygenase).6Arabidopsis thaliana.. The genetically engineered bacteria of claim 5 , wherein the cyp79B2 claim 5 , cyp71a13 claim 5 , and cyp79B3 are from7bacteriumbacterium. The of any of - claim 5 , wherein the comprises a gene or gene cassette for producing indole-3-propionic acid.8bacterium. The genetically engineered bacteria of claim 7 , wherein the engineered comprises gene sequence encoding tryptophan ammonia lyase.9Rubrivivax benzoatilyticus.. The genetically engineered bacyteris of claim 8 , wherein the tryptophan ammonia lyase is from10bacterium. The genetically engineered of any of - claim 8 , wherein the engineered bacterium comprises one or more gene sequences encoding indole-3-acrylate reductase.11bacteriumClostridum botulinum.. The genetically engineered of claim 10 , wherein the ndole-3-acrylate reductase is from12bacteriumbacterium. The genetically engineered of any of - claim 10 , wherein the engineered comprises gene ...

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Номер записи: 38
09-01-2020 дата публикации

SACCHAROMYCES CEREVISIAE STRAINS

Номер: US20200010793A1
Автор: Bonander Nicklas
Принадлежит:

The present invention relates to a method of preparing a strain of sugar fermenting with capability to ferment xylose, wherein said method comprises different procedural steps. The method comprises mating a first sporulated strain with a second haploid strain. Thereafter, screening for mated cells is performed, growing such mated cells, and verifying that mated cells exhibit basic morphology by microscopic inspection. Thereafter, creation of a mixture of the mated cells is performed, subjecting the mixture to continuous chemostat lignocellulose cultivation and obtaining the sugar fermenting cells with capability to ferment xylose is performed. The invention also comprises strains obtained by said method. 1Saccharomyces cerevisiae. A method of preparing a strain of comprising at least one native XKS1 gene in its genome encoding xylulokinase , at least one native XDH1 gene in its genome encoding xylitol dehydrogenase , and at least one modGre3 gene in its genome , said modGre3 gene encoding an amino acid sequence of SEQ ID NO 1 having xylose reductase activity or encoding a fragment of said amino acid sequence having xylose reductase activity , wherein said method comprises the following steps:{'i': 'Saccharomyces cerevisiae', 'a) sporulating a first strain of for providing at least 20 tetrads of said strain,'}{'i': Scheffersomyces stipitis', 'Saccharomyces cerevisiae', 'Saccharomyces cerevisiae,, 'b) introducing DNA, encoding for xylose reductase and xylitol dehydrogenase obtained from and xylulokinase obtained from , into a second strain of'}{'i': Saccharomyces cerevisiae', 'Saccharomyces cerevisiae', 'Saccharomyces cerevisiae, 'c) mating the first sporulated strain with the second strain evolved on xylose and in a haploid state by mixing cells of said haploid strain with each tetrad obtained in step a) to provide mated cells on an YPD agar plate,'}d) screening for mated cells on xylose and geneticin agar plates,e) growing mated cells from step d) in minimal defined ...

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Номер записи: 39
19-01-2017 дата публикации

FLUOROCYCLOPENTENYLCYTOSINE METHODS OF USE

Номер: US20170014411A1
Автор: Kim Deog Joong, Lee Young Bok, Mazhari Reza, Peters Godefridus J., Sarkisjan Dzjemma
Принадлежит: Rexahn Pharmaceuticals, Inc.

The disclosed subject matter provides methods using and kits comprising a compound of formula (I) 2. The method of claim 1 , wherein the dosage is about 500-700 mg/day.3. The method of claim 1 , wherein the dosage is about 6-12 mg/kg/day.4. The method of claim 1 , wherein the oral dosage form is administered 5 to 7 days per week.5. The method of claim 1 , wherein the oral dosage form is administered 5 to 7 days per week for 4 consecutive weeks or for 3 consecutive weeks followed by 1 off-week during which the oral dosage form is not administered.6. The method of claim 5 , wherein a dosing cycle consists of either 3 consecutive weeks of treatment followed by 1 off-week claim 5 , or 4 consecutive weeks of treatment claim 5 , and the oral dosage form is administered for up to 12 dosing cycles.7. The method of claim 1 , wherein the oral dosage form provides a Cof about 700-1 claim 1 ,100 ng/mL after a single administration.8. The method of claim 1 , wherein the oral dosage form provides an AUC(0-24 hours) of about 8 claim 1 ,000-10 claim 1 ,000 hr·ng/mL after a single administration.9. The method of claims 1 , wherein the tumor is pancreatic claims 1 , bladder or colorectal cancer.10. The method of claim 1 , further comprising administering radiation or an anti-tumor agent to the subject.11. The method of claim 1 , further comprising administering an anti-tumor agent selected from the group consisting of antimetabolites claim 1 , DNA-fragmenting agents claim 1 , DNA-crosslinking agents claim 1 , intercalating agents claim 1 , protein synthesis inhibitors claim 1 , topoisomerase I poisons claim 1 , topoisomerase II poisons claim 1 , microtubule-directed agents claim 1 , kinase inhibitors claim 1 , polyphenols claim 1 , hormones claim 1 , hormone antagonists claim 1 , death receptor agonists claim 1 , immune checkpoint inhibitors claim 1 , anti-programmed cell death 1 (PD-1) receptor antibodies and anti-programmed cell death ligand 1 (PD-L1) antibodies.12. The method of ...

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Номер записи: 40
15-01-2015 дата публикации

Methods for treating cardiac conditions

Номер: US20150017255A1
Автор: Hubbard Frank Burrows, III, Thomas J. Koob
Принадлежит: Mimedx Group Inc

Described herein are compositions and methods of treating a cardiac condition using modified placental tissue or an extract of a placental tissue, capable of recruiting stem cells or promoting healing in vivo and in vitro.

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Номер записи: 41
17-01-2019 дата публикации

Novel Peptides and Combination of Peptides for Use In Immunotherapy Against Various Cancers

Номер: US20190015492A1
Автор: Fritsche Jens, MAHR Andrea, SCHOOR Oliver, SINGH Harpreet, SONG Colette, WEINSCHENK Toni
Принадлежит:

The present invention relates to peptides, proteins, nucleic acids and cells for use in immunotherapeutic methods. In particular, the present invention relates to the immunotherapy of cancer. The present invention furthermore relates to tumor-associated T-cell peptide epitopes, alone or in combination with other tumor-associated peptides that can for example serve as active pharmaceutical ingredients of vaccine compositions that stimulate anti-tumor immune responses, or to stimulate T cells ex vivo and transfer into patients. Peptides bound to molecules of the major histocompatibility complex (MHC), or peptides as such, can also be targets of antibodies, soluble T-cell receptors, and other binding molecules. 1. A method of eliciting an immune response in a patient who has cancer , comprising administering to said patient a population of activated T cells that selectively recognize cells , which present a peptide consisting of the amino acid sequence of RLLGTEFQV (SEQ ID NO: 154) , wherein said cancer is selected from the group consisting of glioblastoma , breast cancer , colorectal cancer , renal cell carcinoma , chronic lymphocytic leukemia , hepatocellular carcinoma , non-small cell and small cell lung cancer , non-Hodgkin lymphoma , acute myeloid leukemia , ovarian cancer , pancreatic cancer , prostate cancer , esophageal cancer including cancer of the gastric-esophageal junction , gallbladder cancer and cholangiocarcinoma , melanoma , gastric cancer , urinary bladder cancer , head-and neck squamous cell carcinoma , and uterine cancer.2. The method of claim 1 , wherein the T cells are autologous to the patient.3. The method of claim 1 , wherein the T cells are obtained from a healthy donor.4. The method of claim 1 , wherein the T cells are obtained from tumor infiltrating lymphocytes or peripheral blood mononuclear cells.5. The method of claim 1 , wherein the activated T cells are expanded in vitro.6. The method of claim 1 , wherein the peptide is in a complex ...

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Номер записи: 42
19-01-2017 дата публикации

ENHANCING MICROBIAL METABOLISM OF C5 ORGANIC CARBON

Номер: US20170015988A1
Автор: Armenta Roberto E., Benjamin Jeremy, Merkx-Jacques Alexandra, Muise Denise, Rasmussen Holly, Scaife Mark, Woodhall David
Принадлежит: Mara Renewables Corporation

Provided herein are recombinant microorganisms having two or more copies of a nucleic acid sequence encoding xylose isomerase, wherein the nucleic acid encoding the xylose isomerase is an exogenous nucleic acid. Optionally, the recombinant microorganisms include at least one nucleic acid sequence encoding a xylulose kinase and/or at least one nucleic acid sequence encoding a xylose transporter. The provided recombinant microorganisms are capable of growing on xylose as a carbon source. 1. A recombinant microorganism comprising two or more copies of a nucleic acid sequence encoding xylose isomerase , wherein the nucleic acid encoding xylose isomerase is an exogenous nucleic acid.2. The recombinant microorganism of claim 1 , further comprising at least one nucleic acid sequence encoding a xylulose kinase.3. The recombinant microorganism of claim 2 , further comprising at least one nucleic acid sequence encoding a xylose transporter.4. The recombinant microorganism of claim 1 , wherein the microorganism comprises at least 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 claim 1 , 6 claim 1 , 7 claim 1 , 8 claim 1 , 9 claim 1 , 10 claim 1 , 11 claim 1 , 12 claim 1 , 13 claim 1 , 14 claim 1 , 15 claim 1 , 16 claim 1 , 17 claim 1 , 18 claim 1 , 19 claim 1 , 20 claim 1 , 21 claim 1 , 22 claim 1 , 23 claim 1 , 24 claim 1 , 25 claim 1 , 26 claim 1 , 27 claim 1 , 28 claim 1 , 29 claim 1 , 30 claim 1 , 31 claim 1 , 32 claim 1 , 33 claim 1 , 34 claim 1 , 35 claim 1 , 36 claim 1 , 37 claim 1 , 38 claim 1 , 39 claim 1 , or 40 copies of the exogenous nucleic acid sequence encoding xylose isomerase.5. The recombinant microorganism of claim 1 , wherein the 6 nucleic acid sequence encoding the xylose isomerase is at least 90% identical to SEQ ID NO:2.6. The recombinant microorganism of claim 2 , wherein the microorganism comprises at least 2 claim 2 , 3 claim 2 , 4 claim 2 , 5 claim 2 , 6 claim 2 , 7 claim 2 , 8 claim 2 , 9 claim 2 , 10 claim 2 , 11 claim 2 , 12 claim 2 , 13 claim 2 , 14 claim ...

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Номер записи: 43
19-01-2017 дата публикации

METHODS AND STRAINS FOR PRODUCING BIOPRODUCTS IN AUREOBASIDIUM PULLULANS

Номер: US20170016039A1
Автор: LEATHERS Timothy D., PRICE Neil P., Skory Christopher D.
Принадлежит:

The present disclosure provides methods for producing bioproducts from novel genetically altered strains of . Methods and materials for the construction of these strains, examination of the bioproducts and analysis and isolation of the bioproducts from genetically altered strains is provided. Genetically altered strains in which one or more genes encoding biosynthetic enzymes are knocked out is detailed and the benefits of using such strains described. 1. A method of producing arabitol-liamocins , comprising the steps of{'i': 'A. pullulans', 'a) growing a culture of comprising a biologically pure strain lacking a functional MPD1 gene under conditions sufficient to support the production of arabitol-liamocins, wherein said conditions comprise the substantial absence of arabitol; and'}b) collecting said arabitol-liamocins from at least part of said culture, thereby producing arabitol-liamocins.2. The method of claim 1 , wherein the biologically pure strain comprises NRRL 67079.3. The method of claim 1 , wherein said conditions further comprise a growth medium containing glucose as the sole carbon source.4. A method of producing one or more bioproducts claim 1 , comprising the steps of{'i': 'A. pullulans', 'a) growing a culture of comprising a biologically pure strain lacking a functional MDH2 gene and lacking a functional MPD1 gene under conditions sufficient to support the production of one or more bioproducts selected from the group consisting of a liamocin and an exophilin; and'}b) collecting said one or more bioproducts from at least part of said culture, thereby producing the bioproduct.5. The method of claim 4 , wherein said conditions comprise a growth medium containing glucose or fructose as the sole carbon source.6. The method of claim 4 , wherein the bioproduct is an exophilin.7. The method of claim 4 , wherein the bioproduct is a liamocin and wherein said liamocin has a head group comprising lactose claim 4 , glucose claim 4 , mannose claim 4 , galactose ...

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Номер записи: 44
21-01-2016 дата публикации

COMPOSITIONS AND METHODS FOR BIOLOGICAL PRODUCTION OF ISOPRENE

Номер: US20160017374A1
Автор: Leonard Effendi, Minshull Jeremy, Ness Jon Edward, Purcell Thomas Joseph
Принадлежит:

The present disclosure provides compositions and methods for biologically producing isoprene using methanotrophic bacteria that utilize carbon feedstock, such as methane or natural gas.

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Номер записи: 45
18-01-2018 дата публикации

A Process for the Preparation of Nucleic Acid by Means of 3'-O-Azidomethyl Nucleotide Triphosphate

Номер: US20180016609A1
Автор: Gordon R. MCINROY, Jiahao HUANG, Michael C. Chen, Radu A. LAZAR
Принадлежит: Nuclera Nucleics Ltd

The invention relates to a method of nucleic acid synthesis comprising the use of 3′-O-azidomethyl blocked nucleotide triphosphates which comprises the step of adding a capping group to any uncleaved 3′-O-azidomethyl groups and to the use of kits comprising said capping groups in a method of nucleic acid synthesis. The invention also relates to capped nucleotide triphosphates and 3′-O-azidomethyl capping groups.

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Номер записи: 46
18-01-2018 дата публикации

PIK3CA NOVEL MUTATIONS DETECTION FOR DIAGNOSIS OF ACQUIRED CETUXIMAB RESISTANCE IN METASTATIC COLORECTAL CANCER PATIENTS

Номер: US20180016640A1
Автор: Wang Yan
Принадлежит:

Disclosed is a kit for detecting drug resistance of cetuximab in the treatment of metastatic colorectal cancer. The kit comprises a substance used for detecting gene mutations in Exon 19 of the PIK3CA gene, and may further comprise a specification recording the following contents: if Exon 19 in the PIK3CA gene of a patient with metastatic colorectal cancer as a subject to be tested, who is intended to receive cetuximab treatment or is receiving cetuximab treatment and does not have drug resistance, has at least one of K944N, F930S, V955G, V955I, and K966E mutations, the subject to be tested will develop drug resistance or will be a candidate to develop drug resistance when receiving or continuing to receive cetuximab for treating metastatic colorectal cancer. 1. A kit for detecting or assisting in detecting acquired cetuximab resistance in metastatic colorectal cancer , comprising a substance used for detecting whether Exon 19 in a PIK3CA gene has a gene mutation wherein Exon 19 is SEQ ID NO: 1.2. The kit according to claim 1 , wherein the kit further comprises a specification recording the following contents:if the Exon 19 in PIK3CA gene of a patient with metastatic colorectal cancer as a subject to be tested, who is intended to receive cetuximab treatment or is receiving cetuximab treatment and does not develop drug resistance, has at least one of following (a)-(e) mutations, the subject to be tested will develop drug resistance or will be a candidate that develops drug resistance when receiving or continuing to receive cetuximab for treating metastatic colorectal cancer:(a) Lysine at the 944th amino acid of the PIK3CA gene coding protein mutates into Asparagine;(b) Phenylalanine at 930th amino acid of the PIK3CA gene coding protein mutates into Serine;(c) Valine at the 955th amino acid of the PIK3CA gene coding protein mutates into Isoleucine;(d) Valine at the 955th amino acid of the PIK3CA gene coding protein mutates into Glycine; and(e) Lysine at the 966th ...

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Номер записи: 47
17-01-2019 дата публикации

MULTIMODAL TRAIL MOLECULES AND USES IN CELLULAR THERAPIES

Номер: US20190016767A1
Автор: Shah Khalid
Принадлежит: The General Hospital Corporation

Described herein are novel compositions comprising multimodal TRAIL agents and cells engineered to express such multimodal TRAIL agents, including cells encapsulated in a scaffold or matrix, for use in the treatment of disorders such as cancer. 122.-. (canceled)23. A composition comprising a T cell comprising a nucleic acid encoding a polypeptide comprising a soluble TRAIL fusion protein comprising a reporter module , a linker module , and a therapeutic TRAIL module comprising an extracellular domain of human TRAIL of SEQ ID NO: 1.24. The composition of claim 23 , wherein the cell is encapsulated in a matrix or scaffold.25. The composition of claim 23 , wherein the therapeutic TRAIL module comprises amino acids 39-281 of SEQ ID NO: 1.26. The composition of claim 23 , wherein the therapeutic TRAIL module comprises amino acids 95-281 of SEQ ID NO: 1.27. The composition of claim 23 , wherein the therapeutic TRAIL module comprises amino acids 114-281 of SEQ ID NO: 1.28. The composition of claim 23 , wherein the therapeutic TRAIL module consists of amino acids 114-281 of SEQ ID NO: 1.29. The composition of claim 24 , wherein the matrix comprises a synthetic extracellular matrix.30. The composition of claim 24 , wherein the matrix is biodegradable.31. The composition of claim 29 , wherein the synthetic extracellular matrix comprises a thiol-modified hyaluronic acid and a thiol-reactive cross-linker molecule.32. The composition of claim 30 , wherein the thiol-reactive cross-linker molecule is polyethylene glycol diacrylate.33. The composition of claim 23 , further comprising a nucleic acid sequence encoding HSV-TK.34. The composition of claim 23 , wherein the polypeptide comprises claim 23 , in the following N-terminal to C-terminal order claim 23 , a reporter module claim 23 , a linker module of at least 8 amino acids claim 23 , and a therapeutic TRAIL module comprising an extracellular domain of human TRAIL of SEQ ID NO: 1.35. The composition of claim 34 , wherein the ...

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Номер записи: 48
17-01-2019 дата публикации

OLIGONUCLEOTIDE COMPOSITIONS WITH ENHANCED EFFICIENCY

Номер: US20190017051A1
Автор: Taylor Margaret, Woolf Tod
Принадлежит:

The oligonucleotide compositions of the present invention make use of combinations of oligonucleotides. In one aspect, the invention features an oligonucleotide composition including at least 2 different oligonucleotides targeted to a target gene. This invention also provides methods of inhibiting protein synthesis in a cell and methods of identifying oligonucleotide compositions that inhibit synthesis of a protein in a cell. 133.-. (canceled)34. A composition comprising a combination of double-stranded oligonucleotides , the combination consisting of three or four different double-stranded oligonucleotides ,wherein each double-stranded oligonucleotide is targeted to a different sequence within a single target gene,each double-stranded oligonucleotide consists of two separate strands,each strand is between 15 and 40 nucleomonomers in length,wherein the combination is capable of RNA interference (RNAi).35. The composition of wherein each strand is between 20 and 30 nucleomonomers in length.36. The composition of wherein at least one of the oligonucleotides comprises at least one modified sugar moiety or at least one modified internucleoside linkage.37. The composition of wherein each double-stranded oligonucleotide comprises a nucleotide sequence that is at least 90% identical to a portion of the target gene.38. The composition of wherein at least one of the double-stranded oligonucleotides is covalently linked to an agent to facilitate cellular uptake.39. The composition of claim 34 , wherein the target gene is a mammalian gene.40. A composition comprising a combination of double-stranded oligonucleotides claim 34 , the combination consisting of two different double-stranded oligonucleotides claim 34 ,wherein each double-stranded oligonucleotide is targeted to a different sequence within a single target gene,each double-stranded oligonucleotide consists of two separate strands,each strand is between 15 and 40 nucleomonomers in length,wherein the combination is ...

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Номер записи: 49
17-01-2019 дата публикации

METHODS, CELLS AND REAGENTS FOR PRODUCTION OF ISOPRENE, DERIVATIVES AND INTERMEDIATES THEREOF

Номер: US20190017076A1
Автор: Conradie Alex Van Eck
Принадлежит:

This application describes methods, including non-naturally occurring methods, for biosynthesizing 3-hydroxy-3-methylglutaryl-coA and intermediates thereof, as well as non-naturally occurring hosts for producing 3-hydroxy-3-methylglutaryl-coA. This application also describes methods, including non-naturally occurring methods, for biosynthesizing isoprene and intermediates thereof, as well as non-naturally occurring hosts for producing isoprene. 112-. (canceled)13. A non-naturally occurring host capable of producing 3-hydroxy-3-methylglutaryl-CoA , said host comprising:(a) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.7.7 or EC 1.2.1.- enzyme; or at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.2.1.39 or EC 1.2.1.5 enzyme;', 'at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.2.1.2. enzyme; or (c) both (a) and (b); and, '(b) at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1.74 or EC 4.1.1.43 enzyme;'}at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.3.8.4 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 6.4.1.4 enzyme; andat least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.2.1.18 enzyme.14. The host of claim 13 , wherein said host is capable of producing isoprene and comprises:at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 1.1.1.34 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 2.7.1.36 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 2.7.4.2 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 4.1.1.33 enzyme;at least one exogenous nucleic acid encoding a polypeptide having the activity of an EC 5.3.3.2 ...

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Номер записи: 50
17-01-2019 дата публикации

ENZYMATIC PRODUCTION OF D-TAGATOSE

Номер: US20190017083A1
Автор: WICHELECKI Daniel Joseph
Принадлежит: BONUMOSE LLC

The current disclosure provides a process for enzymatically converting a saccharide into tagatose. The invention also relates to a process for preparing tagatose where the process involves converting fructose 6-phosphate (F6P) to tagatose 6-phosphate (T6P), catalyzed by an epimerase, and converting the T6P to tagatose, catalyzed by a phosphatase. 1. An enzymatic process for preparing tagatose , the process comprising the steps of:(i) converting fructose 6-phosphate (F6P) to tagatose 6-phosphate (T6P) catalyzed by a fructose 6-phosphate epimerase (F6PE); and(ii) converting the T6P produced to tagatose catalyzed by a tagatose 6-phosphate phosphatase (T6PP);wherein steps (i)-(ii) are conducted in a single reaction vessel.213-. (canceled)14. The process of claim 1 , wherein the F6PE comprises an amino acid sequence having at least 90% claim 1 , at least 95% claim 1 , at least 97% claim 1 , at least 99% claim 1 , or 100% sequence identity with SEQ ID NOS.: 2 claim 1 , 4 claim 1 , 6 claim 1 , 8 claim 1 , or 11.1516-. (canceled)17. The process of claim 1 , wherein the T6PP comprises an amino acid sequence having at least 90% claim 1 , at least 95% claim 1 , at least 97% claim 1 , at least 99% claim 1 , or 100% sequence identity with SEQ ID NOS.: 13 claim 1 , 15 claim 1 , or 17.1821-. (canceled)22. The process of claim 14 , wherein the T6PP comprises an amino acid sequence having at least 90% claim 14 , at least 95% claim 14 , at least 97% claim 14 , at least 99% claim 14 , or 100% sequence identity with SEQ ID NOS.: 13 claim 14 , 15 claim 14 , or 17. This application claims priority to U.S. application Ser. No. 15/743,481, filed Jan. 10, 2018; which claims priority to PCT International Application No. PCT/US2016/054838 filed Sep. 30, 2016; and to U.S. Provisional Application No. 62/236,226, filed Oct. 2, 2015, which is incorporated herein by reference.The invention relates to preparation of the sugar D-tagatose. More specifically, the invention relates to methods of ...

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Номер записи: 51
21-01-2021 дата публикации

Methods for controlling gene expression

Номер: US20210017514A1
Автор: Alexander Morgan Jones, Bo Larsen
Принадлежит: University of Cambridge

The invention relates to methods for precisely controlling expressions of a target gene in an organism using a light-inducible kinase and a response regulator. The invention also relates to nucleic acid constructs and nucleic acids encoding the light-inducible kinase and response regulator, as well as organisms expressing these constructs.

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Номер записи: 52
26-01-2017 дата публикации

STEM CELLS FOR ANTI-ANGIOGENIC THERAPY IN AGE-RELATED MACULAR DEGENERATION, DIABETIC RETINOPATHY, CORNEAL VASCULARISATION AND CANCER

Номер: US20170020958A1
Автор: YONG Then Khong
Принадлежит:

The present invention relates to production of a stem cell expressing an anti-angiogenic protein. The stem cells are used to inhibit angiogenesis for treatment of macular degeneration, corneal vascularisation, cancer and diabetic retinopathy. 1. Genetically-engineered mesenchymal stem cells (MSCs) having a recombinant vector carrying a vascular endothelial growth factor receptor (VEGFR) gene and expressing a vascular endothelial growth factor receptor (VEGFR) polypeptide , wherein the said stem cells inhibit angiogenesis in human body.2. The stem cells as claimed in claim 1 , wherein the VEGFR gene is VEGFR1.3. The stem cells as claimed in claim 1 , wherein the said VEGFR polypeptide is a soluble form of VEGFR.4. The stem cells as claimed in claim 1 , wherein the VEGFR polypeptide is a human FLT-1 protein.5. The stem cells as claimed in claim 1 , wherein the recombinant vector is a plasmid or a viral vector.6. The stem cells as claimed in claim 5 , wherein the plasmid vector is pBLAST-hsFLT-1.7. The stem cells as claimed in claim 1 , wherein the mesenchymal stem cells are isolated from umbilical cord.8. The stem cells as claimed in claim 6 , wherein the plasmid vector is transfected into the stem cells by cationic lipid transfection.9. The stem cells as claimed in claim 1 , wherein the stem cells express proteins having an amino acid sequence of SEQ ID NO.1.10. The stem cells as claimed in claim 9 , wherein the stem cells express proteins having sequence 50 to 100% homology to SEQ ID NO.1.11. The stem cells as claimed in claim 1 , wherein the angiogenesis is inhibited in patients having disease or disorder selected from a group comprising of macular degeneration claim 1 , cancer claim 1 , diabetic retinopathy claim 1 , lymphangiogenesis claim 1 , retinal neovascularisation claim 1 , thyroid hyperplasia claim 1 , preeclampsia claim 1 , rheumatoid arthritis and osteo-arthritis claim 1 , Alzheimer's disease claim 1 , obesity claim 1 , pleural effusion claim 1 , ...

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Номер записи: 53
25-01-2018 дата публикации

FLUOROCYCLOPENTENYLCYTOSINE METHODS OF USE

Номер: US20180021338A1
Автор: Kim Deog Joong, Lee Young Bok, Mazhari Reza, Peters Godefridus J., Sarkisjan Dzjemma
Принадлежит: Rexahn Pharmaceuticals, Inc.

The disclosed subject matter provides methods using and kits comprising a compound of formula (I) 2. The method of claim 1 , further comprising administering the subject with the compound of formula (I) if an increased expression of UCK2 is measured in the tumor cell or tissue.3. The method of claim 2 , wherein the increased expression level is determined when the amount of UCK2 expression is greater than a predetermined level of UCK2 in a non-tumor cell.4. The method of claim 3 , wherein the non-tumor cell is a non-tumor cell of the subject.5. The method of claim 1 , wherein the increased expression level is by 5% or more.6. The method of claim 1 , wherein the tumor cell is ovarian cancer; metastatic breast cancer; adenocarcinoma of the pancreas; gastrointestinal cancer such as colorectal adenocarcinoma or cancer of the esophagus claim 1 , stomach claim 1 , pancreas claim 1 , small bowel claim 1 , hepatobiliary tract claim 1 , colon claim 1 , rectum or anus; bladder cancer such as metastatic bladder cancer claim 1 , muscle invasive bladder cancer or non-muscle invasive bladder cancer; cervical cancer; lung cancer; non-small cell lung cancer; or renal cell carcinoma.7. The method of claim 1 , wherein the tumor cell is lung cancer cell.8. The method of claim 1 , wherein the tumor cell is non-small cell lung cancer cell.10. The method of claim 9 , wherein contacting the tumor cell or tissue with a compound of formula (I) is accomplished by administering the compound of formula (I) to the subject.11. The method of claim 9 , wherein measuring one of protein kinases or p53 expression level in the tumor cell or tissue after contact with the compound of formula (I) is conducted by collecting a second sample of tumor cell or tissue from the subject and measuring one of protein kinases or p53 expression level in the second sample.12. The method of claim 9 , further comprising administering the subject with the compound of formula (I) if an increase in one of protein kinases ...

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Номер записи: 54
26-01-2017 дата публикации

METHODS TO INCREASE PHOTOSYNTHETIC RATES IN PLANTS

Номер: US20170022512A1
Автор: Altpeter Fredy, Jaikumar Nikhil S., Karan Ratna, Long Stephen P., Moose Stephen P., Swaminathan Kankshita, XIE Liang
Принадлежит:

Disclosed herein are transgenic plants and plant cells having increased photosynthetic rate, increased biomass production, and/or improved cold tolerance compared to control plants (such as non-transgenic plants of the same species as the transgenic plants). In some examples, the transgenic plants/plant cells contain a plant transformation vector including a nucleic acid encoding a pyruvate orthophosphate dikinase (PPDK) polypeptide. Also disclosed herein are methods for making the transgenic plants, for instance by introducing into progenitor cells of the plant a plant transformation vector including a nucleic acid that encodes a PPDK polypeptide, and growing the transformed progenitor cells to produce a transgenic plant, in which the PPDK nucleic acid is expressed. Further disclosed herein are PPDK-encoding nucleic acids, PPDK polypeptides, and plant transformation vectors of use in producing the transgenic plants or plant cells. 1. A transgenic C4 or CAM plant comprising a plant transformation vector comprising a heterologous nucleic acid encoding a pyruvate orthophosphate dikinase (PPDK) polypeptide:having an amino acid sequence (1) at least 90% identical to SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, or (2) comprising the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12; andwherein the transgenic plant expresses an increased amount of PPDK nucleic acid or PPDK protein compared to a control plant.2. The transgenic plant of claim 1 , wherein the heterologous nucleic acid comprises a nucleic acid sequence (1) at least 80% identical to SEQ ID NO: 3 claim 1 , SEQ ID NO: 5 claim 1 , SEQ ID NO: 7 claim 1 , SEQ ID NO: 9 claim 1 , SEQ ID NO: 11 claim 1 , or (2) comprising the nucleic acid sequence of SEQ ID NO: 3 claim 1 , SEQ ID NO: 5 claim 1 , SEQ ID NO: 7 claim 1 , SEQ ID NO: 9 claim 1 , SEQ ID NO: 11 claim 1 , or (3) comprising positions 35533 . . . 23640 of SEQ ID NO: 1 and a PPDK cDNA ...

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Номер записи: 55
25-01-2018 дата публикации

MICROORGANISMS AND METHODS FOR THE CO-PRODUCTION OF ETHYLENE GLYCOL AND THREE CARBON COMPOUNDS

Номер: US20180023101A1
Автор: KOCH Daniel Johannes, Lopes Mateus Schreiner, Parizzi Lucas Pedersen, Zeidler Ane Fernanda Beraldi
Принадлежит:

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG) and one or more three-carbon compounds such as acetone, isopropanol or propene. The MEG and one or more three-carbon compounds described herein are useful as starting material for production of other compounds or as end products for industrial and household use. The application further relates to recombinant microorganisms co-expressing a C2 branch pathway and a C3 branch pathway for the production of MEG and one or more three-carbon compounds. Also provided are methods of producing MEG and one or more three-carbon compounds using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally the products MEG and one or more three-carbon compounds. 2. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises at least one endogenous or exogenous nucleic acid molecule encoding a secondary alcohol dehydrogenase that catalyzes the conversion of acetone to isopropanol.3. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises: at least one endogenous or exogenous nucleic acid molecule encoding a secondary alcohol dehydrogenase that catalyzes the conversion of acetone to isopropanol; and at least one endogenous or exogenous nucleic acid molecule encoding a dehydratase that catalyzes the conversion of isopropanol to propene.4. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises one or more modifications selected from the group consisting of:(a) a deletion, insertion, or loss of function mutation in a gene encoding a D-xylose isomerase that catalyzes the conversion of D-xylose to D-xylulose;(b) a deletion, insertion, or loss of function mutation in a gene encoding a glycolaldehyde dehydrogenase that catalyzes the conversion of glycolaldehyde to glycolic acid; and(c) a deletion, ...

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Номер записи: 56
25-01-2018 дата публикации

NOVEL USE

Номер: US20180023108A1
Автор: CHEN Michael C., HUANG Jiahao, LAZAR Radu A., MCINROY Gordon R.
Принадлежит:

The invention relates to the use of specific terminal deoxynucleotidyl transferase (TdT) enzymes in a method of nucleic acid synthesis, to methods of synthesizing nucleic acids, and to the use of kits comprising said enzymes in a method of nucleic acid synthesis. The invention also relates to the use of terminal deoxynucleotidyl transferases and 3′-blocked nucleotide triphosphates in a method of template independent nucleic acid synthesis. 1. A method of nucleic acid synthesis comprising a step of providing a terminal deoxynucleotidyl transferase (TdT) enzyme comprising an amino acid sequence having at least 95% sequence identity to any one of SEQ ID NOs: 1 to 5 and 8 , such as any of SEQ ID NOs: 1 , 2 , or 8.2. (canceled)3. A method of nucleic acid synthesis , comprising steps of:(a) providing an initiator sequence;{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(b) adding a 3′-blocked nucleotide triphosphate to said initiator sequence in the presence of a terminal deoxynucleotidyl transferase (TdT) as defined in ;'}(c) removal of all reagents from the initiator sequence;(d) cleaving the blocking group from the 3′-blocked nucleotide triphosphate in the presence of a cleaving agent; and(e) removal of the cleaving agent.4. The method as defined in claim 3 , wherein greater than 1 nucleotide is added by repeating steps (b) to (e).5. The method as defined in claim 3 , wherein the 3′-blocked nucleotide triphosphate is blocked by either a 3′-O-azidomethyl claim 3 , 3′-aminoxy or 3′-O-allyl group.6Saccharomyces cerevisiae.. The method as defined in claim 3 , wherein the terminal deoxynucleotidyl transferase (TdT) is added in the presence of an extension solution comprising one or more buffers claim 3 , such as Tris or cacodylate claim 3 , one or more salts; and/or inorganic pyrophosphatase claim 3 , such as purified claim 3 , recombinant inorganic pyrophosphatase from7. The method as defined in claim 3 , wherein step (b) is performed at a pH range of between 5 and 10 ...

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Номер записи: 57
10-02-2022 дата публикации

Method for rescuing influenza virus and composition therefor

Номер: US20220041997A1
Автор: Dai Dongsheng, Li Xun, Tian Xin
Принадлежит:

The present invention relates to a new method for rescuing an influenza virus and a composition therefor. The method comprises providing a host cell stably integrated with and expressing influenza virus PA, PB1, PB2 and NP genes, and introducing an influenza virus rescue system in which a stop codon is introduced into the PA, PB1, PB2 and NP genes respectively into the host cell to achieve virus rescue. The produced virus particles can be used as a live attenuated influenza vaccine, which is characterized in that, since the genes encoding the related proteins are mutated, it has no replication and proliferation ability in human and normal animal cells, and replication and proliferation can be achieved only in the host cells constructed above and it can fully stimulate the body immunity and effectively protect the body while ensuring the safety. 1. A method for rescuing an influenza virus , characterized in that a mammalian host cell stably expressing influenza virus PA , PB1 , PB2 and NP genes is provided , and an influenza virus rescue system comprising mutant PA , PB1 , PB2 and NP genes is introduced into the aforementioned host cell to achieve rescue , wherein the mutations make the influenza virus rescue system unable to rescue intact virus in natural mammalian cells.2. The method according to claim 1 , wherein the method comprises the following steps:(1) constructing a single- or multiple-plasmid system encoding the PA, PB1, PB2 and NP genes;(2) introducing the single- or multiple-plasmid system of step (1) into a mammalian cell, and screening a host cell stably expressing the four genes;(3) constructing recombinant plasmids for the mutant PA, PB1, PB2 and NP genes and recombinant plasmids encoding the four genes HA, NA, M and NS respectively to form an influenza virus rescue system, the mutations are achieved by introducing a TAG codon into each of the four gene sequences;(4) co-transfecting the influenza virus rescue system constructed in step (3) into the ...

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Номер записи: 58
25-01-2018 дата публикации

LATERAL FLOW DEVICE AND METHOD OF USE

Номер: US20180024129A1
Автор: Strong William
Принадлежит:

Lateral flow devices, methods and kits for performing lateral flow western blot assays are provided. 1. A lateral flow device comprising: 'wherein the wicking pad has a first end, a second end and two lateral edges;', 'a wicking pad composed of a porous material, the wicking pad having a region for applying a substrate comprising immobilized analytes; and'}a first reservoir comprising a stack of a plurality of reagent layers located on the first end of the wicking pad; anda second reservoir comprising an absorbent pad located on the second end of the wicking pad.2. The device of claim 1 , wherein each of the plurality of reagent layers comprises a reagent immobilized in an absorbent pad.3. The device of claim 2 , wherein each of the plurality of reagent layers has a different reagent therein.4. The device of claim 2 , wherein the reagent is selected from the group consisting of a primary antibody claim 2 , a secondary antibody claim 2 , a first wash solution claim 2 , and a second wash solution.5. The device of claim 1 , wherein the plurality of reagent layers claim 1 , starting at a reagent layer in contact with the wicking pad claim 1 , comprises a first reagent layer having a primary antibody claim 1 , a second reagent layer having a first wash solution claim 1 , a third reagent layer having a secondary antibody claim 1 , and a fourth reagent layer having a second wash solution.6. The device of claim 5 , wherein the fourth reagent layer is at least twice the thickness of the third reagent layer.7. The device of claim 5 , further comprising a fifth reagent layer comprising the second wash solution.8. The device of claim 5 , wherein the volume of the second wash solution is at least twice the volume of the secondary antibody.9. The device of claim 5 , wherein each of the plurality of reagent layers is formed of an absorbent pad and at least a portion of the first reagent layer is in intimate contact with the wicking pad.10. The device of claim 1 , wherein the ...

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Номер записи: 59
28-01-2021 дата публикации

MLK-Regulated microRNAs in Angiogenesis and Tumor Development

Номер: US20210023118A1
Автор: John F. Keaney, JR., Roger J. Davis, Shashi Kant, Siobhan CRAIGE
Принадлежит: University of Massachusetts UMass

Methods for treating cancer, and for reducing angiogenesis in a tissue, comprising administering one or more of a miR-371 oligonucleotide; a miR-146 oligonucleotide; or an inhibitor of MLK2/3 activity.

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Номер записи: 60
24-01-2019 дата публикации

METHODS FOR EXTENDING THE REPLICATIVE CAPACITY OF SOMATIC CELLS DURING AN EX VIVO CULTIVATION PROCESS

Номер: US20190024079A1
Автор: DESMET Danielle Nicole, GENOVESE Nicholas J., SCHULZE Eric
Принадлежит:

A product and process for extending the replicative capacity of metazoan somatic cells using targeted genetic amendments to abrogate inhibition of cell-cycle progression during replicative senescence and derive clonal cell lines for scalable applications and industrial production of metazoan cell biomass. An insertion or deletion mutation using guide RNAs targeting the first exon of the transcript encoding each protein is created using CRISPR/Cas9. Targeted amendments result in inactivation of p15 and p16 proteins which increases the proliferative capacity of the modified cell populations relative to their unaltered parental populations. Combining these amendments with ancillary telomerase activity from a genetic construct directing expression of a telomerase protein homolog from a TERT gene, increases the replicative capacity of the modified cell populations indefinitely. One application is to manufacture skeletal muscle for dietary consumption using cells from the poultry species ; another is from the livestock species 1. A process for extending the replicative capacity of a metazoan somatic cell population comprising:decoupling retinoblastoma protein inhibition of cell division cycle advancement during replicative senescence by abrogating cyclin-dependent kinase inhibitor (“CKI”)-mediated stabilization of a retinoblastoma protein using genetic amendment;maintaining telomerase activity by transducing the metazoan somatic cell population with a genetic construct (SEQ ID NO 11) directing ectopic expression of functional telomere reverse transcriptase (“TERT”) protein;maintaining a bank of cells that is a master cell bank having the genetic amendment and ectopic expression of TERT protein;cultivating cells from the master cell bank in an ex vivo milieu that is a cultivated cell biomass; andharvesting the cultivated cell biomass for dietary consumption.2. The process of claim 1 , wherein the genetic amendment comprises inactivating a p15 protein by genetic amendment ...

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Номер записи: 61
23-01-2020 дата публикации

IMPROVED GLYCEROL FREE ETHANOL PRODUCTION

Номер: US20200024619A1
Автор: DE BRUIJN Hans Marinus Charles Johannes, DE WAAL Paulus Petrus, SCHMITZ Jozef Petrus Johannes
Принадлежит:

The invention relates to a recombinant cell, preferably a yeast cell comprising one or more genes coding for an enzyme having glycerol dehydrogenase activity, one or more genes coding dihydroxyacetone kinase (E.C. 2.7.1.28 and/or E.C. 2.7.1.29); one or more genes coding for an enzyme in an acetyl-CoA-production pathway and one or more genes coding for an enzyme having at least NAD dependent acetylating acetaldehyde dehydrogenase activity (EC 1.2.1.10 or EC 1.1.1.2), and optionally one or more genes coding for a glycerol transporter. This cell can be used for the production of ethanol and advantageously produces little or no glycerol. 1. A recombinant cell , optionally a yeast cell , said recombinant cell comprising:one or more genes coding for an enzyme having glycerol dehydrogenase activity;one or more genes coding dihydroxyacetone kinase (E.C. 2.7.1.28 and/or E.C. 2.7.1.29);one or more genes coding for an enzyme in an acetyl-CoA-production pathway; and{'sup': '+', 'one or more genes coding for an enzyme having at least NAD dependent acetylating acetaldehyde dehydrogenase activity (EC 1.2.1.10 or EC 1.1.1.2); and optionally'}one or more genes coding for a glycerol transporter.2. The Cell according to wherein the enzyme having glycerol dehydrogenase activity is a NAD linked glycerol dehydrogenase (EC 1.1.1.6).3. The Cell according to wherein the enzyme having glycerol dehydrogenase activity is a NADP linked glycerol dehydrogenase (EC 1.1.1.72).4. The recombinant cell according to wherein the one or more genes coding for an enzyme in an acetyl-CoA-production pathway comprises:one or more genes coding for an enzyme having phosphoketolase (PKL) activity (EC 4.1.2.9 or EC 4.1.2.22) or an enzyme having an amino acid sequence according SEQ ID NO: 5, 6, 7, or 8, or functional homologues thereof having a sequence identity of at least 50%, and/orone or more genes coding for an enzyme having phosphotransacetylase (PTA) activity (EC 2.3.1.8) or an enzyme having an amino acid ...

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Номер записи: 62
23-01-2020 дата публикации

Composition for producing tagatose and method of producing tagatose using the same

Номер: US20200024627A1
Автор: Chan Hyoung Lee, Eun Jung Choi, Hyun Kug Cho, II Hyang PARK, Seong Bo Kim, Sungjae YANG, Young Mi Lee
Принадлежит: CJ CHEILJEDANG CORP

Provided are a composition for producing tagatose, comprising fructose-4-epimerase, and a method of producing tagatose using the same.

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Номер записи: 63
28-01-2021 дата публикации

Cell-Free Expression System Having Novel Inorganic Polyphosphate-Based Energy Regeneration

Номер: US20210024912A1
Автор: Humbert Michael, Koglin Alexander
Принадлежит:

The invention relates to an in vitro cell-free expression system incorporating a novel inorganic polyphosphate-based energy regeneration system. In certain embodiments, the invention includes a cell-free expression system where the cellular energy source, ATP, is regenerated from inorganic polyphosphate using a dual enzyme system. In this embodiment, this dual enzyme system may include thermostable Adenosyl Kinase, and/or Polyphosphate Kinase enzymes. 1. A cell-free protein expression system comprising:a bioreactor having:{'i': Geobacillus', 'Geobacillus, 'claim-text': knock-out expression of OmpT-homologue;', 'knock-out expression of RNaseI;', 'knock-out expression of DNA-methylation dependent DNase;', 'reduce expression of culture density-dependent sporulation operon;', 'overexpress sigma factor RpoD; and', 'overexpress RNA polymerase (RNAP)., 'a reaction mixture containing a thermostable cell extract having all cell-free reaction components necessary for in vitro macromolecule synthesis and wherein said is genetically modified to include at least one of the followingat least one nucleic acid synthesis template; a quantity of purified Adenosyl Kinase (AdK) enzyme; and', 'a quantity of purified Polyphosphate Kinase (PPK) enzyme;', 'a quantity of inorganic polyphosphate (PPi); and', 'a quantity of adenosine monophosphate (AMP)., 'a cellular adenosine triphosphate (ATP) energy regeneration system comprisingwherein said AdK and PPK enzymes work synergistically to regenerate cellular ATP energy from PPi and AMP.2GeobacillusGeobacillus stearothermophilus.. The system of claim 1 , wherein said comprises3. The system of claim 1 , wherein said cell-free reaction components comprises cell-free reaction components selected from the group consisting of:amino acids;polyphosphate;Tris-Acetate;{'sub': '2', 'Mg(OAc);'}{'sup': '+', 'K-glutamate;'}amino-acetate;NaCl;KCl;{'sub': '2', 'MgCl;'}DTT;octyl-b-glycoside;NAD;NADP;sorbitol;FADH;ATP;GTP, UTP, and/or CTP;CoA;PLP; andSAM.4. The ...

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Номер записи: 64
28-01-2021 дата публикации

GENETICALLY ENGINEERED MICROORGANISMS AND PROCESSES FOR THE PRODUCTION OF CANNABINOIDS FROM A CARBON SOURCE PRECURSOR

Номер: US20210024968A1
Автор: CHAN Brian, DAVIDOVICH Adam, Gupta Neil, Ladha Alim
Принадлежит:

A method is provided for biosynthetic production of cannabinoids in microorganisms from a carbon source precursor. This method describes the genetic modifications needed to engineer microorganisms to produce cannabinoids as well as a method for identifying and quantifying cannabinoids from fermentation broth. A system is also provided for tuning the method to produce different cannabinoids of interest by systematically modulating the enzymes encoded by the genetic modifications introduced in the microorganism. 1. A method for producing at least one cannabinoid from a carbon source precursor , comprising:genetically modifying a microorganism to express enzymes for converting the carbon source precursor into at least one cannabinoid within the genetically modified bacterial strain.2. The method according to claim 1 , wherein the carbon source precursor is glucose and the method further comprises converting the glucose to hexanoate.3. The method according to claim 2 , wherein the at least one cannabinoid comprises cannabigerolic acid.4E. coli.. The method according to claim 3 , wherein the microorganism is a bacterial strain5. The method according to claim 4 , wherein genetically modifying the bacterial strain comprises recombinantly incorporating a mutated FadD gene at the genomic location of a FadE gene of the bacterial strain to express the mutated FadD enzyme and simultaneously knock out the FadE gene of the bacterial strain.6. The method according to claim 5 , wherein the mutated FadD gene comprises a nucleotide sequence of SEQ ID NO: 10.7. The method according to claim 4 , wherein genetically modifying the bacterial strain comprises transforming the bacterial strain to express olivetol synthase claim 4 , olivetolic acid cyclase claim 4 , and CsPT1.8. The method according to claim 7 , wherein the olivetol synthase comprises a first amino acid sequence comprising the amino acid sequence of SEQ ID NO: 2 claim 7 , wherein the olivetolic acid cyclase comprises a ...

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Номер записи: 65
28-01-2021 дата публикации

BIOMARKER COMPOSITION FOR DIAGNOSING RADIATION-RESISTANT CANCER OR FOR PREDICTING PROGNOSIS OF RADIATION THERAPY CONTAINING PMVK AS ACTIVE INGREDIENT

Номер: US20210025007A1
Автор: Choi Eun Kyung, JEONG Seong-Yun, JU Eun Jin, KO Eun Jung, PARK Seok Soon, PARK Yun-Yong, SONG Si Yeol
Принадлежит: ENHANCEDBIO INC.

The present invention relates to a biomarker composition for diagnosing radiation-resistant cancer comprising PMVK as an active ingredient and a method of diagnosing radiation-resistant cancer using the same, and when the PMVK is knocked down, it is confirmed that the survival rate of cancer cells decreases during radiation treatment, and based on this, the possibility as a factor related to radiation therapy resistance to cancer was suggested, and the PMVK can be used as a new target to enhance the effect of radiation therapy on human cancer cells. 1. A method for diagnosing radiation-resistant cancer , comprising:(1) measuring mRNA expression level of a PMVK gene or expression level of a PMVK protein from a sample isolated from a cancer patient;(2) comparing the mRNA expression level of the PMVK gene or the expression level of the PMVK protein with a control sample; and(3) determining that it is radiation-resistant cancer when the mRNA expression level of the PMVK gene or the expression level of the PMVK protein is higher than that of the control sample.2. The method of claim 1 , wherein the cancer is lung cancer or pancreatic cancer.312-. (canceled)13. A method for enhancing radiation sensitivity to cancer cells comprising a step of administering to a subject in need thereof a pharmaceutical composition comprising a phosphomevalonate kinase (PMVK) protein expression inhibitor or activity inhibitor as an active ingredient.14. The method of claim 13 , wherein the PMVK protein expression inhibitor is any one selected from the group consisting of antisense nucleotides claim 13 , small interfering RNA (siRNA) and short hairpin RNA (shRNA) claim 13 , that complementarily bind to mRNA of a PMVK gene.15. The method of claim 13 , wherein the PMVK protein activity inhibitor is any one selected from the group consisting of low molecular compounds claim 13 , peptides claim 13 , peptide mimetics claim 13 , aptamers claim 13 , antibodies and natural products claim 13 , that ...

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Номер записи: 66
29-01-2015 дата публикации

CELLS GENETICALLY MODIFIED TO COMPRISE PANCREATIC ISLET GLUCOKINASE AND USES THEREOF

Номер: US20150030574A1
Автор: Simpson Ann Margaret, Tao Chang
Принадлежит: UNIVERSITY OF TECHNOLOGY, SYDNEY

The present invention relates generally to a population of cells genetically modified to produce insulin in a glucose responsive manner and uses thereof. More particularly, the present invention relates to a population of cells genetically modified to produce insulin in response to physiologically relevant levels of glucose and uses thereof. The cells of the present invention are useful in a wide variety of applications, in particular in the context of therapeutic and prophylactic regimes directed to the treatment of diabetes and/or the amelioration of symptoms associated with diabetes, based on the transplantation of the cells of the present invention into mammals requiring treatment. Also facilitated is the design of in vitro based screening systems for testing the therapeutic effectiveness and/or toxicity of potential adjunctive treatment regimes. 1. A genetically modified mammalian hepatocyte , which hepatocyte secretes insulin , said comprising a transfected nucleic acid molecule encoding a human pancreatic islet glucokinase.2. The genetically modified cell of wherein said mammalian cell is a human cell.3. The genetically modified hepatocyte of claim 1 , wherein said hepatocyte is transfected with a nucleic acid molecule encoding insulin thereof.4. The genetically modified hepatocyte of wherein said hepatocyte is an Huh7ins cell.5. The genetically modified hepatocyte of claim 1 , wherein said hepatocyte expresses glucokinase in response to an extracellular glucose concentration in the range of 3-8 mM.6. The genetically modified hepatocyte of wherein said hepatocyte expresses glucokinase in response to an extracellular glucose concentration in the range of 3.5-7 mM.7. The genetically modified hepatocyte of wherein said hepatocyte expresses glucokinase in response to an extracellular glucose concentration in the range of 4-6 mM.8. The genetically modified hepatocyte of wherein said hepatocyte expresses glucokinase in response to an extracellular glucose ...

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Номер записи: 67
29-01-2015 дата публикации

MODULATING THE INTERACTION BETWEEN ZO-2/TJP2 AND A SNAIL ZINC FINGER TRANSCRIPTION FACTOR FAMILY MEMBER

Номер: US20150031748A1
Автор: Goh Choon Peng, Hunziker Walter
Принадлежит: AGENCY FOR SCIENCE, TECHNOLOGY AND RESEARCH

There is provided a method of identifying candidate agents capable of modulating interaction between a first polypeptide and a second polypeptide, wherein the first polypeptide is ZO-2/TJP2 or a functional variant thereof and the second polypeptide is a Snail zinc finger transcription factor family member or a functional variant thereof. 1. A method of identifying candidate agents capable of modulating interaction between a first polypeptide and a second polypeptide , wherein the first polypeptide is ZO-2/TJP2 or a functional variant thereof and the second polypeptide is a Snail zinc finger transcription factor family member or a functional variant thereof , the method comprising:a. contacting the first polypeptide with the second polypeptide and a candidate agent; andb. determining whether the binding of the first polypeptide with the second polypeptide is decreased or increased in the presence of said candidate agent when compared with a control.2. The method of claim 1 , wherein the ZO-2/TJP2 is selected from the group consisting of SEQ ID NO: 1 claim 1 , SEQ ID NO: 2 claim 1 , SEQ ID NO: 3 claim 1 , SEQ ID NO: 4 claim 1 , SEQ ID NO: 5 claim 1 , SEQ ID NO: 6 claim 1 , SEQ ID NO: 7 claim 1 , SEQ ID NO: 8 and SEQ ID NO: 9.3. The method of claim 1 , wherein the Snail zinc finger transcription factor family member is selected from the group consisting of SNAI1 (Snail) claim 1 , SNAI2 (Slug) claim 1 , SNAI3 (Smuc) claim 1 , Scratch 1 and Scratch 2.4. The method of claim 3 , wherein the Snail zinc finger transcription factor family member is selected from the group consisting of SEQ ID NO: 10 claim 3 , SEQ ID NO: 11 claim 3 , SEQ ID NO: 12 claim 3 , SEQ ID NO: 13 claim 3 , SEQ ID NO: 14 claim 3 , SEQ ID NO: 15 claim 3 , SEQ ID NO: 16 claim 3 , SEQ ID NO: 17 claim 3 , SEQ ID NO: 18 and SEQ ID NO: 19.5. The method of claim 1 , wherein the functional variant of ZO-2/TJP2 has at least 80% sequence identity with said ZO-2/TJP2.6. The method of claim 3 , wherein the ...

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Номер записи: 68
31-01-2019 дата публикации

NOVEL FUSIONS AND METHOD FOR DETECTING SAME

Номер: US20190033293A1
Автор: Fujita Naoya, KATAYAMA Ryohei, Sakata Seiji, Takeuchi Kengo, Togashi Yuki
Принадлежит:

It is intended to reveal a polynucleotide serving as a novel causative gene of a cancer and, on the basis of this finding, to provide a method for detecting the polynucleotide or a polypeptide encoded thereby, a kit and a primer set for the detection, a method for screening for a substance that inhibits the polypeptide, and a pharmaceutical composition for the treatment of a cancer, containing the inhibiting substance. The detection method of the present invention detects a BRAF fusion protein or a fusion gene encoding the fusion protein, or a PXN or GMDS fusion protein or a fusion gene encoding the fusion protein in a digestive organ-derived sample obtained from a subject. 127-. (canceled)28. A method for screening for a substance that inhibits an activity and/or expression of a polypeptide , comprising the steps of:(1) contacting the polypeptide or a cell expressing the polypeptide with a test substance;(2) analyzing whether or not to inhibit the activity and/or expression of the polypeptide; and(3) selecting the substance that inhibits the activity and/or expression of the polypeptide,wherein the polypeptide is selected from the group consisting of the following polypeptides (a) to (d):(a) a polypeptide consisting of the amino acid sequence represented by SEQ ID NO: 2 (PXN-BRAF) or SEQ ID NO: 4 (GMDS-BRAF);(b) a polypeptide comprising the amino acid sequence represented by SEQ ID NO: 2 or SEQ ID NO: 4, and having tumorigenicity;(c) a polypeptide comprising an amino acid sequence with 80% or higher identity to the amino acid sequence represented by SEQ ID NO: 2 or SEQ ID NO: 4, and having tumorigenicity; and(d) a polypeptide comprising an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 2 or SEQ ID NO: 4 by the deletion, substitution, and/or insertion of one or several amino acids, and having tumorigenicity.29. The screening method according to claim 28 , wherein the substance that inhibits the activity and/or expression of the ...

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Номер записи: 69
08-02-2018 дата публикации

Methods for enhancing the delivery of molecules across the blood brain barrier

Номер: US20180036420A1
Автор: Henri Lichenstein, Jonathan M. Rothberg, Tian Xu
Принадлежит: LAM Therapeutics Inc

The present invention relates to methods and compositions for enhancing the delivery of an agent across the blood brain barrier utilizing a composition comprising an inhibitor of PIKfyve.

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Номер записи: 70
11-02-2016 дата публикации

Microorganisms of corynebacterium which can utilize xylose and method for producing l-lysine using same

Номер: US20160040200A1
Автор: Chang Gyeom Kim, Hyung Joon Kim, Jin Seok Sung, Jun Ok Moon, Kwang Ho Lee, Kyung Han Lee, Lan Huh, So Yeon Rah
Принадлежит: CJ CHEILJEDANG CORP

The present invention relates to microorganisms of corynebacterium which can utilize xylose and to a method for producing L-lysine using same. More particularly, the present invention relates to microorganisms of corynebacterium which are modified, in which genes encoding xylose isomerase and xylulokinase which are xylose synthases are introduced to express the xylose synthase. The present invention also relates to a method for producing L-lysine, comprising a step of culturing the modified microorganisms of corynebacterium using xylose as a carbon source, and recovering L-lysine from the culture.

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Номер записи: 71
09-02-2017 дата публикации

SACCHAROMYCES CEREVISIAE STRAINS

Номер: US20170037361A1
Автор: Bonander Nicklas
Принадлежит:

The present invention relates to a method of preparing a strain of sugar fermenting with capability to ferment xylose, wherein said method comprises different procedural steps. The method comprises mating a first sporulated strain with a second haploid strain. Thereafter, screening for mated cells is performed, growing such mated cells, and verifying that mated cells exhibit basic morphology by microscopic inspection. Thereafter, creation of a mixture of the mated cells is performed, subjecting the mixture to continuous chemostat lignocellulose cultivation and obtaining the sugar fermenting cells with capability to ferment xylose is performed. The invention also comprises strains obtained by said method. 1Saccharomyces cerevisiae. A strain of comprising at least one native XKS1 gene in its genome encoding xylulokinase , at least one native XDH1 gene in its genome encoding xylitol dehydrogenase , and at least one modGre3 gene in its genome , said modGre3 gene encoding an amino acid sequence of SEQ ID NO 1 having xylose reductase activity or encoding a fragment of said amino acid sequence having xylose reductase activity , wherein said strain is obtained by the following steps:{'i': 'Saccharomyces cerevisiae', 'a) sporulating a first strain of for providing at least 20 tetrads of said strain,'}{'i': Scheffersomyces stipitis', 'Saccharomyces cerevisiae', 'Saccharomyces cerevisiae,, 'b) introducing DNA, encoding for xylose reductase and xylitol dehydrogenase obtained from and xylulokinase obtained from , into a second strain of'}{'i': Saccharomyces cerevisiae', 'Saccharomyces cerevisiae', 'Saccharomyces cerevisiae, 'c) mating the first sporulated strain with the second strain evolved on xylose and in a haploid state by mixing cells of said haploid strain with each tetrad obtained in step a) to provide mated cells on an YPD agar plate,'}d) screening for mated cells on xylose and geneticin agar plates,e) growing mated cells from step d) in minimal defined xylose liquid ...

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Номер записи: 72
08-02-2018 дата публикации

Method for Secretory Production of Protein

Номер: US20180037918A1
Автор: Itaya Hiroshi, Ito Yumi, Kashima Yukari, Matsuda Yoshihiko, Tsurui Noriko, YAMADA Naoko
Принадлежит: AJINOMOTO CO., INC.

A novel technique for improving secretory production of a heterologous protein by coryneform bacteria is described, as well as a method for secretory production of a heterologous protein. A coryneform bacterium is described that has been modified to have a specific mutation so as to harbor a phoS gene, and when cultured, is able to to produce a heterologous protein by secretory production. 1. A method for producing a heterologous protein comprising:culturing a coryneform bacterium having a genetic construct that allows for secretory expression of a heterologous protein; andcollecting the heterologous protein produced by secretory production,wherein the coryneform bacterium has been modified so as to harbor a phoS gene encoding PhoS protein with a mutation,wherein the mutation results in improved secretory production of the heterologous protein as compared to a bacterium without the mutation,wherein the genetic construct comprises, in the direction from 5′ to 3′, a promoter sequence that is able to function in the coryneform bacterium, a nucleic acid sequence encoding a signal peptide that is able to function in the coryneform bacterium, and a nucleic acid sequence encoding the heterologous protein, andwherein the heterologous protein is expressed as a fusion protein with the signal peptide.2. The method according to claim 1 , wherein the mutation is replacement of an amino acid residue other than a histidine residue that is autophosphorylated with another amino acid residue in a wild-type PhoS protein.3. The method according to claim 1 , wherein the mutation is replacing an autophosphorylated amino acid residue that is not a histidine residue in a HisKA domain of a wild-type PhoS protein with another amino acid residue.4. The method according to claim 1 , wherein the mutation is replacing an amino acid residue at position 302 in SEQ ID NO: 4 with an amino acid residue other than an aromatic amino acid residue and a histidine residue in a wild-type PhoS protein.5. A ...

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Номер записи: 73
08-02-2018 дата публикации

FUNGAL STRAINS AND METHODS OF USE

Номер: US20180037919A1
Автор: Bodie Elizabeth A., NGUYEN Rochelle, RABINOVICH Roman, SWEIGARD James A., Virag Aleksandra, Ward Michael
Принадлежит: DANISCO US INC.

Provided are improved fungal strains and use thereof, wherein the fungal strains are capable of producing an altered level of proteins, enzymes, variants and other substances of interest. 1. An engineered fungal strain , capable of producing an altered level of a protein of interest as compared to a parental strain , wherein the engineered fungal strain comprises a variant histidine kinase gene.2. An engineered fungal strain , capable of producing an altered level of a protein of interest as compared to a parental strain , wherein the engineered fungal strain comprises a mutation that causes altered sensitivity or resistance to external osmotic pressure as compared to the parental strain.3. An engineered fungal strain , capable of producing an altered level of a protein of interest as compared to a parental strain , wherein the engineered fungal strain comprises a mutation that causes altered sensitivity or resistance to a fungicide as compared to the parental strain.49-. (canceled)10. The engineered fungal strain of claim 1 , wherein the parental strain is an Ascomycete fungal strain.1112-. (canceled)13. A transformed fungal strain or a derivative fungal strain thereof capable of producing an altered level of a protein of interest as compared to a parental strain claim 1 , wherein the transformed fungal strain or the derivative fungal strain comprises a variant histidine kinase gene or a mutation that causes altered sensitivity or resistance to external osmotic pressure as compared to the parental strain.1419-. (canceled)20. The fungal strain of claim 13 , wherein the parental strain is an Ascomycete fungal strain.2129-. (canceled)3023. The method of claim claim 13 , wherein the parental strain is an Ascomycete fungal strain.3132-. (canceled)33. A method of producing a protein of interest comprising fermenting the engineered fungal strain of claim 1 , wherein the engineered claim 1 , transformed or derivative fungal strain secretes the protein of interest claim 1 , ...

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Номер записи: 74
12-02-2015 дата публикации

PRACTICAL METHOD FOR ENZYMATICALLY SYNTHESIZING CYCLIC DI-GMP

Номер: US20150044724A1
Автор: Ishige Kazuya, Tanabe Kaori
Принадлежит:

A practical method for enzymatically synthesizing c-di-GMP with excellent productivity is provided. A diguanylate cyclase having physical and chemical characteristics (A) to (F): (A) catalytic action on reaction “2 GTP→c-di-GMP”; (B) a molecular weight of 19800±2000; (C) an optimum pH of 7.3 to 9.4; (D) an optimum temperature of 35 to 60° C.; (E) thermal stability as the remaining activity of 90% or higher after heated for 60 minutes under conditions of 50° C. and pH 7.8; and (F) the presence of GGDEF domain and the lack of amino acid sequence KXXD in the i-site. 1. A diguanylate cyclase with physical and chemical characteristics of:(A) catalytic action on reaction “2 GTP→c-di-GMP”;(B) a molecular weight of 19800±2000;(C) an optimum pH of 7.3 to 9.4;(D) an optimum temperature of 35 to 60° C.;(E) thermal stability as the remaining activity of 90% or higher after heated for 60 minutes under conditions of 50° C. and pH 7.8; and(F) the presence of GGDEF domain and the lack of amino acid sequence KXXD in the i-site.2. The diguanylate cyclase according to claim 1 , wherein the diguanylate cyclase has one or more amino acid sequences selected from the group consisting of:(G) the amino acid sequence shown under SEQ ID NO:6,(H) an amino acid sequence having identity of 90% or higher with the amino acid sequence shown under SEQ ID NO:6, having 54th residue glycine in the amino acid sequence shown under SEQ ID NO:6 conserved, and with the lack of amino acid sequence KXXD in the i-site,(I) an amino acid sequence equivalent to the amino acid sequence shown under SEQ ID NO:6 including deletion, substitution, insertion, or addition of one or several amino acids, having 54th residue glycine in the amino acid sequence shown under SEQ ID NO:6 conserved, and with the lack of amino acid sequence KXXD in the i-site, and(J) an amino acid sequence coded by the base sequence of a nucleic acid that hybridizes, under stringent conditions, with a nucleic acid having a base sequence ...

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Номер записи: 75
07-02-2019 дата публикации

ENGINEERED CYANOBACTERIUM AND ITS APPLICATION FOR PRODUCING ACETATE

Номер: US20190040424A1
Автор: Lu Wei, Shen Roa-Pu
Принадлежит:

The present invention provides an engineered cyanobacterium, comprising at least one plasmid selected from three novel pathways to produce acetate, which can convert atmospheric carbon dioxide as a raw material into acetate. The present invention also constructs the expression plasmid for three different transporters specific to acetate to be expressed in cyanobacteria, which comprises putative ABC transporter (AatA), succinate/acetate: proton symporter (SatP) and acetate/glycolate: cation symporter (ActP). Therefore, the engineered cyanobacteria of the present invention can produce 0.58 mg/L to 3.54 mg/L of acetate per hour. 1. An engineered cyanobacterium , comprising at least one plasmid selected from the group consisting of:(1) a plasmid containing pyruvate decarboxylase gene (pdc), and acetaldehyde dehydrogenase B gene (aldB) or 3-hydroxypropionaldehyde dehydrogenase gene (aldH);(2) a plasmid containing phosphate acetyltransferase gene (pta) or phosphate acetyltransferase gene (eutD), and acetate kinase gene (ackA); and {'i': 'Bifidobacterium strains.', 'wherein the plasmid is incorporated into a host cyanobacterium chromosome, and fpk is obtained from'}, '(3) a plasmid containing acetate kinase gene (ackA), fructose-1,6-biphosphatase gene (fbp) and fructose-6-phosphoketolase gene (fpk),'}2Synechococcus elongates. The engineered cyanobacterium of claim 1 , wherein the host cyanobacterium is sp. PCC 7942.3. The engineered cyanobacterium of claim 1 , wherein the plasmid further contains a transporter gene.4. The engineered cyanobacterium of claim 3 , wherein the transporter gene is putative ABC transporter gene (aatA) claim 3 , succinate/acetate: proton symporter gene (satP) or acetate/glycolate: cation symporter gene (actP).5Escherichia coli.. The engineered cyanobacterium of claim 1 , wherein the pta claim 1 , the eutD claim 1 , the aldB and the aldH are obtained from6Zymomonas mobilis.. The engineered cyanobacterium of claim 1 , wherein the pdc is obtained ...

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Номер записи: 76
07-02-2019 дата публикации

RECOMBINANT MICROORGANISM OF GENUS KOMAGATAEIBACTER, METHOD OF PRODUCING CELLULOSE BY USING THE SAME, AND METHOD OF PRODUCING THE MICROORGANISM

Номер: US20190040476A1
Автор: Chung Soonchun, LEE Sunghaeng, Park Jinhwan
Принадлежит:

Provided is a microorganism of the genus having enhanced cellulose productivity and yield, a method of producing cellulose by using the same, and a method of producing the microorganism. 1Komagataeibacter. A recombinant microorganism of the genus comprising a genetic modification that increases expression or activity of polyphosphate kinase (PPK).2. The microorganism of claim 1 , wherein the genetic modification increases expression of a gene encoding the polyphosphate kinase.3. The microorganism of claim 1 , wherein the genetic modification is an increase in the copy number of a gene encoding the polyphosphate kinase or a modification of an expression regulatory sequence of the gene encoding the polyphosphate kinase.4. The microorganism of claim 1 , wherein the polyphosphate kinase belongs to EC 2.7.4.1.5. The microorganism of claim 1 , wherein the polyphosphate kinase catalyzes both the forward and reverse reaction of converting NTP+ (phosphate)n to NDP+ (phosphate)n+1 claim 1 , and has higher catalytic activity for the reverse reaction than for the forward reaction.6. The microorganism of claim 5 , wherein the polyphosphate kinase has higher catalytic activity for conversion of NDP to NTP in a reaction using GDP claim 5 , CDP claim 5 , or UDP as a substrate claim 5 , compared to using ADP.7. The microorganism of claim 1 , wherein the polyphosphate kinase is a polypeptide having a sequence identity of about 85% or more with an amino acid sequence of SEQ ID NO: 44 claim 1 , 46 claim 1 , or 48.8Rhodobacterales. The microorganism of claim 1 , wherein the polyphosphate kinase is a Silicibacter polyphosphate kinase or a polyphosphate kinase.9. The microorganism of claim 1 , wherein the microorganism has enhanced cellulose productivity as compared to a microorganism of the same type without the genetic modification that increases expression or activity of the polyphosphate kinase (PPK).10. The microorganism of claim 1 , further comprising a genetic modification that ...

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Номер записи: 77
16-02-2017 дата публикации

PRECISION GENE TARGETING TO A PARTICULAR LOCUS IN MAIZE

Номер: US20170044558A1
Автор: Ainley W. Michael, BING James W., Corbin David R., EVANS STEVEN L., Petolino Joseph F., Sastry-Dent Lakshmi, Thompson Steven A., Webb Steven R., WELTER Mary E., ZHOU Ning
Принадлежит:

The present invention claims methods for the stable integration of exogenous DNA into a specific locus, E32, in the maize genome through the use of zinc finger nucleases. Maize plants and plant parts that were transformed by the methods of the invention are claimed. The invention is useful for creating desirable traits such as herbicide resistance, herbicide tolerance, insect resistance, insect tolerance, disease resistance, disease tolerance, stress tolerance, and stress resistance in maize The E32 locus represents a superior site for inserting foreign genes because native agronomic phenotypes are not disturbed. 1. A method for integrating one or more exogenous nucleic acid sequences into the genome of a maize cell having an E32 locus , said method comprising: making a double-stranded cleavage in the E32 locus using a site specific nuclease , ligating into the cleavage a polynucleotide comprising the one or more exogenous sequences into the genome of the maize cell within the E32 locus.2. The method of wherein the site specific nuclease comprises one or more zinc finger nuclease selected from the group shown in Table 1A3. The method of claim 1 , further comprising the step of expressing a product of the one or more exogenous sequences.4. The method of claim 2 , further comprising the step of expressing a product of the one or more exogenous sequences.5. The method of claim 1 , wherein the one or more exogenous nucleic acid sequences comprise a coding sequence claim 1 , a regulatory sequence claim 1 , or a target site for a DNA-binding domain.6. The method of claim 2 , wherein the one or more exogenous nucleic acid sequences comprise a coding sequence claim 2 , a regulatory sequence claim 2 , or a target site for a DNA-binding domain.7. The method of claim 3 , wherein the one or more exogenous nucleic acid sequences comprise a coding sequence claim 3 , a regulatory sequence claim 3 , or a target site for a DNA-binding domain.8. The method of claim 4 , wherein the ...

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Номер записи: 78
03-03-2022 дата публикации

ANAEROBIC FERMENTATIVE PRODUCTION OF FURANDICARBOXYLIC ACID

Номер: US20220064683A1
Автор: GOUVEA Iuri Estrada, Marchesin Mariana Trovo, ROMÃO DUMARESQ Aline Silva, SIMOES Marcos Rogerio
Принадлежит:

The present disclosure provides recombinant microorganisms and methods for the anaerobic production of 2,4-furandicarboxylic acid from one or more carbon sources. The microorganisms and methods provide redox-balanced and ATP positive pathways for co-producing 2,4-furandicarboxylic acid with ethanol and for co-producing 2,4-furandicarboxylic acid with ethanol and 1-propanol. The method provides recombinant microorganisms that express endogenous and/or exogenous nucleic acid molecules encoding polypeptides that catalyze the conversion of a carbon source into 2,4-furandicarboxylic acid and that coupled the 2,4-furandicarboxylic acid pathway with an additional metabolic pathway.

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Номер записи: 79
03-03-2022 дата публикации

Biosynthetic methods and systems for producing monosaccharides

Номер: US20220064685A1
Автор: Tahereh Karimi
Принадлежит: Cemvita Factory Inc

The present disclosure is related to biosynthetic methods of forming monosaccharides, and systems for generating the same. A benefit of the methods and systems disclosed herein can include the sustainable production of monosaccharides in an automated process. A benefit of the methods and systems herein can be the generation of monosaccharides from renewable source materials. An additional benefit of the methods and systems herein can include the use of abundant feedstocks, such as carbon dioxide, for the efficient generation of select monosaccharides for use as nutrients and for other useful applications. Another benefit of the methods and systems disclosed herein can include reduction of excess carbon dioxide from the environment.

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Номер записи: 80
03-03-2022 дата публикации

CELL-FREE PRODUCTION OF RIBONUCLEIC ACID

Номер: US20220064688A1
Автор: Abshire James Robbins, Blake William Jeremy, Cunningham Drew S., Gupta Mehak, MacEachran Daniel
Принадлежит: GreenLight Biosciences, Inc.

Provided herein, in some aspects, are methods and compositions for cell-free production of ribonucleic acid.

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Номер записи: 81
03-03-2022 дата публикации

METHOD FOR REDUCING MISINCORPORATION OF NON-CANONICAL BRANCHED-CHAIN AMINO ACIDS

Номер: US20220064692A1
Автор: CORCOLES GARCIA Angel, HAUPTMANN Peter, LATTEMANN Claus Tobias, MATZEN Arne, Neubauer Peter
Принадлежит:

The present invention relates to a method for producing a recombinant polypeptide of interest in a microbial host cell, comprising (a) introducing a polynucleotide encoding the polypeptide of interest into a microbial host cell which has been modified such that an enzymatic activity selected from the group consisting of ketol-acid reductoisomerase (NADP(+)) activity (EC 1.1.1.86), acetohydroxyacid synthase activity (EC 2.2.1.6), aspartate kinase activity (EC 2.7.2.4), homoserine dehydrogenase activity (EC 1.1.1.3), and L-threonine dehydratase activity (EC 4.3.1.19) is modulated in said microbial host cell as compared to the enzymatic activity in an unmodified microbial host cell, and (b) expressing said polypeptide of interest in said microbial host cell. Moreover, the present invention relates to a method for reducing misincorporation of at least one non-canonical branched-chain amino acid into a recombinant polypeptide of interest expressed in a microbial host cell.

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Номер записи: 82
08-05-2014 дата публикации

Microorganisms and methods for production of specific length fatty alcohols and related compounds

Номер: US20140127765A1
Автор: Anthony P. Burgard, Robin E. Osterhout
Принадлежит: Genomatica Inc

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.

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Номер записи: 83
14-02-2019 дата публикации

A BACTERIAL CELL FACTORY FOR EFFICIENT PRODUCTION OF ETHANOL FROM WHEY

Номер: US20190048368A1
Автор: Dantoft Shruti Harnal, Jensen Peter Ruhdal, LIU Jianming, Solem Christian
Принадлежит:

The invention relates to a method for homo-ethanol production from lactose using a genetically modified lactic acid bacterium of the invention, where the cells are provided with a substrate comprising dairy waste supplemented with an amino nitrogen source (such as acid hydrolysed corn steep liquor). The invention further relates to genetically modified lactic acid bacterium and its use for homo-ethanol production from lactose in dairy waste. The lactic acid bacterium comprises both genes (lacABCD, LacEF, lacG) encoding enzymes catalysing the lactose catabolism pathway; and transgenes (pdc and adhB) encoding enzymes catalysing the conversion of pyruvate to ethanol. Additionally a number of genes (ldh, pta and adhE) are deleted in order to maximise homo-ethanol production as compared to production of lactate, acetoin and acetate production. 1. A method for ethanol production using a genetically engineered lactic acid bacterium comprising the steps of:a. introducing a genetically modified lactic acid bacterium into an aqueous culture medium;b. incubating the culture of (a);c. recovering ethanol produced by said culture during step (b), and optionally wherein the aqueous culture medium comprises:', 'I. whey permeate or residual whey permeate, and', wherein the genetically engineered lactic acid bacterium comprises transgenes encoding:', 'i. a polypeptide having pyruvate decarboxylase (PDC) activity (EC 4.1.1.1); and', 'wherein the genome of said lactic acid bacterium comprises genes encoding polypeptides having:', 'ii. a polypeptide having alcohol dehydrogenase B activity (EC 1.1.1.1); and'}, 'iii. lactose-specific phosphotransferase system (PTS) activity (EC 2.7.1.69)', 'iv. phospho-β-D-galactosidase activity (EC 3.2.1.85)', 'v. galactose-6-phosphate isomerase activity (EC 5.3.1.26),', 'vi. D-tagatose-6-phosphate kinase activity (EC 2.7.1.114), and', 'wherein the genome of said lactic acid bacterium is deleted for genes or lacks functional genes or genes encoding ...

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Номер записи: 84
25-02-2021 дата публикации

Microorganism for producing putrescine or ornithine and process for producing putrescine or ornithine using them

Номер: US20210054347A1
Автор: Baek Seok Lee, Hee Kyoung Jung, Hong Xian Li, Hye Won Um, Hyo Hyoung Lee, Kyoung Min Lee, Su Jin Park, Young Lyeol Yang
Принадлежит: CJ CHEILJEDANG CORP

Disclosed is a modified microorganism producing putrescine or ornithine, and a method for producing putrescine or ornithine using the same.

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Номер записи: 85
25-02-2021 дата публикации

REGULATION OF CSR SYSTEM FOR PRODUCTION OF LYSINE AND LYSINE-DERIVED PRODUCTS

Номер: US20210054423A1
Автор: Chen Ling, Chou Howard, LEI Yunfeng, LIU Xiucai
Принадлежит:

The invention provides microorganisms genetically modified to overexpress biofilm dispersal related polypeptides to enhance the production of lysine and lysine derivatives by the microorganism, method of generating such microorganism, and methods of producing lysine and lysine derivatives using the genetically modified microorganisms. 1. A genetically modified host cell comprising an exogenous nucleic acid encoding a CsrB sRNA or a CsrC sRNA , wherein the host cell overexpresses the CsrB sRNA or CsrC sRNA relative to a counterpart host cell that has not been modified to express the exogenous nucleic acid; and has at least one additional genetic modification to increase production of lysine or a lysine derivative compared to a wildtype host cell.2. The genetically modified host cell of claim 1 , wherein:(i) the amino acid derivative is cadaverine;(ii) the endogenous nucleic acid encoding CsrB sRNA comprises a nucleotide sequence having at least 85% identity to SEQ ID NO:16 and the endogenous nucleic acid encoding CsrC sRNA comprises a nucleotide sequence having at least 85% identity to SEQ ID NO:17;(iii) a CsrB or CsrC is heterologous to the host cell;(iv) the host cell overexpresses a lysine decarboxylate; or(v) the host cell overexpresses one or more lysine biosynthesis polypeptides.3. (canceled)4. The genetically modified host cell of claim 1 , wherein the CsrB sRNA comprises the nucleic acid sequence of SEQ ID NO:16 and the CsrC sRNA comprises the nucleic acid sequence of SEQ ID NO:17.5. (canceled)6. The genetically modified host cell of claim 1 , wherein the exogenous nucleic acid encoding the CsrB or CsrC is encoded by an expression vector introduced into the cell claim 1 , wherein the expression vector comprises the exogenous nucleic acid operably linked to a promoter claim 1 , or the exogenous nucleic acid is integrated into the host chromosome.7. (canceled)8. (canceled)9. (canceled)10. The genetically modified host cell of claim 2 , wherein the one or more ...

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Номер записи: 86
13-02-2020 дата публикации

MICROORGANISMS AND METHODS FOR THE CO-PRODUCTION OF ETHYLENE GLYCOL AND ISOBUTENE

Номер: US20200048662A1
Автор: Galzerani Felipe, KOCH Daniel Johannes, Lopes Mateus Schreiner
Принадлежит:

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG) and isobutene. The application further relates to recombinant microorganisms co-expressing a C2 branch pathway and a C3 branch pathway for the production of MEG and isobutene. Also provided are methods of producing MEG and isobutene using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally the products MEG and isobutene. 3. The recombinant microorganism of or , wherein the recombinant microorganism further comprises one or more modifications selected from the group consisting of:(a) a deletion, insertion, or loss of function mutation in a gene encoding a D-xylulose-5-kinase that catalyzes the conversion of D-xylulose to D-xylulose-5-phosphate;(b) a deletion, insertion, or loss of function mutation in a gene encoding a glycolaldehyde dehydrogenase that catalyzes the conversion of glycolaldehyde to glycolic acid; and(c) a deletion, insertion, or loss of function mutation in a gene encoding a lactate dehydrogenase that catalyzes the conversion of pyruvate to lactate.4. The recombinant microorganism of any one of - , wherein an endogenous or exogenous xylose isomerase catalyzes the conversion of D-xylose to D-xylulose.5. The recombinant microorganism of any one of - , wherein the recombinant microorganism further expresses at least one exogenous nucleic acid molecule encoding a xylose reductase or aldose reductase that catalyzes the conversion of D-xylose to xylitol and at least one exogenous nucleic acid molecule encoding a xylitol dehydrogenase that catalyzes the conversion of xylitol to D-xylulose.7. The recombinant microorganism of claim 6 , wherein the recombinant microorganism further comprises one or more modifications selected from the group consisting of:(a) a deletion, insertion, or loss of function mutation in a gene encoding a D-xylose isomerase that catalyzes the conversion of D ...

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Номер записи: 87
13-02-2020 дата публикации

Production method for substance using atp

Номер: US20200048675A1
Автор: Misato MATSUI, Noriyuki Ito, Yoshihiko Yasohara
Принадлежит: Kaneka Corp

A method of producing a substance includes synthesizing a molecule at least by mixing substrates, a synthase, adenosine triphosphate (ATP), a polyphosphate kinase 2, and a polyphosphoric acid mixture. The polyphosphoric acid mixture includes 50% or more of polyphosphoric acid with a degree of polymerization of not less than 15. Adenosine diphosphate (ADP) is generated from the ATP during the synthesis. The synthesis is coupled with an ATP regeneration reaction in which the ATP is regenerated by the polyphosphate kinase 2 from the ADP and the polyphosphoric acid.

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Номер записи: 88
23-02-2017 дата публикации

Method for Producing L-Amino Acids in Corynebacteria Using a Glycine Cleavage System

Номер: US20170051324A1
Автор: Bathe Brigitte, HASSELMEYER Marleen, OCHROMBEL Ines
Принадлежит: EVONIK DEGUSSA GmbH

It has been found, surprisingly, that the strain comprises a very effective glycine cleavage system. 115-. (canceled)16. A glycine cleavage system comprising one or more of the enzymes GcvP , GcvT and GcvH , wherein:a) GcvP comprises a sequence at least 80% identical to the sequence of SEQ ID NO:40;b) GcvT comprises a sequence at least 80% identical to the sequence of SEQ ID NO:42; andc) GcvH comprises a sequence at least 80% identical to the sequence of SEQ ID NO:38.17. The glycine cleavage system of claim 16 , wherein said system comprises at least two of said enzymes.18. The glycine cleavage system of claim 16 , wherein said system comprises all three of said enzymes.19. The glycine cleavage of claim 16 , wherein:a) GcvP comprises a sequence at least 95% identical to the sequence of SEQ ID NO:40;b) GcvT comprises a sequence at least 95% identical to the sequence of SEQ ID NO:42; andc) GcvH comprises a sequence at least 95% identical to the sequence of SEQ ID NO:38.20. The glycine cleavage system of claim 19 , wherein said system comprises all three of said enzymes.21. The glycine cleavage system of claim 16 , wherein said system comprises at least one further polypeptide selected from the group consisting of:a) a LipA enzyme having a sequence at least 80% identical to the sequence of SEQ ID NO:48;b) a LipB enzyme having a sequence at least 80% identical to the sequence of SEQ ID NO:50;c) a Lpd enzyme having a sequence at least 80% identical to the sequence of SEQ ID NO:52;d) a LplA enzyme having a sequence at least 80% identical to the sequence of SEQ ID NO:94;e) a GcvL enzyme having a sequence at least 80% identical to the sequence of SEQ ID NO:96.22. The glycine cleavage system of claim 21 , wherein said system comprises all three of said enzymes and wherein:a) GcvP comprises a sequence at least 95% identical to the sequence of SEQ ID NO:40;b) GcvT comprises a sequence at least 95% identical to the sequence of SEQ ID NO:42; andc) GcvH comprises a sequence at ...

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Номер записи: 89
26-02-2015 дата публикации

Methods for Generating Stably Linked Complexes Composed of Homodimers, Homotetramers or Dimers of Dimers and Uses

Номер: US20150056680A1
Автор: Chang Chien-Hsing, GOLDENBERG David M., McBride William J., Rossi Edmund A.
Принадлежит:

The present invention concerns methods and compositions for stably tethered structures of defined compositions, which may have multiple functionalities and/or binding specificities. Particular embodiments concern homodimers comprising monomers that contain a dimerization and docking domain attached to a precursor. The precursors may be virtually any molecule or structure, such as antibodies, antibody fragments, antibody analogs or mimetics, aptamers, binding peptides, fragments of binding proteins, known ligands for proteins or other molecules, enzymes, detectable labels or tags, therapeutic agents, toxins, pharmaceuticals, cytokines, interleukins, interferons, radioisotopes, proteins, peptides, peptide mimetics, polynucleotides, RNAi, oligosaccharides, natural or synthetic polymeric substances, nanoparticles, quantum dots, organic or inorganic compounds, etc. Other embodiments concern tetramers comprising a first and second homodimer, which may be identical or different. The disclosed methods and compositions provide a facile and general way to obtain homodimers, homotetramers and heterotetramers of virtually any functionality and/or binding specificity. 1. A homodimer , each monomer of the homodimer comprising (i) a dimerization and docking domain (DDD) of human protein kinase A (PKA) regulatory subunit RIIα attached to (ii) an antigen-binding antibody fragment.2. The homodimer of claim 1 , wherein the monomer is a fusion protein.3. The homodimer of claim 1 , wherein the monomer further comprises a linker peptide between the antigen-binding antibody fragment and the DDD.4. The homodimer of claim 1 , wherein the antigen-binding antibody fragment comprises an Fd fragment of an antibody.5. The homodimer of wherein the antigen-binding antibody fragment is selected from the group consisting of a F(ab′) claim 1 , F(ab) claim 1 , Fab claim 1 , Fv claim 1 , scFv and single domain antibody fragment.6. The homodimer of claim 1 , wherein the antigen-binding antibody fragment ...

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Номер записи: 90
01-03-2018 дата публикации

Composition Comprising Neoagarooligosaccharide As Active Ingredient, For Prevention or Treatment of Sepsis or Septic Shock

Номер: US20180055872A1
Автор: HONG Soon Kwang, Hong Sun Joo, Lee Je Hyeon, LEE Moon Hee
Принадлежит:

The present invention relates to a composition for prevention or treatment of sepsis or septic shock, in which the composition includes neoagarooligosaccharide as an active ingredient, and the neoagarooligosaccharide according to the present invention has an excellent effect in terms of immune enhancement by effectively suppressing inflammation, and also exhibits a good effect in preventing sepsis, and therefore can be effectively used in pharmaceuticals and functional foods for prevention or treatment of sepsis or septic shock and immune enhancement. 1. A pharmaceutical composition for prevention or treatment of sepsis or septic shock , the pharmaceutical composition comprising neoagarooligosaccharide , which is prepared from agar or agarose by a DagA enzyme reaction , as an active ingredient.2Streptomyces coelicolor.. The pharmaceutical composition of claim 1 , wherein the DagA enzyme is derived from3. The pharmaceutical composition of claim 2 , wherein the DagA enzyme is represented by amino acid sequences 31 to 309 of SEQ ID NO: 2.4. The pharmaceutical composition of claim 2 , wherein the DagA enzyme is encoded by a nucleotide sequence of SEQ ID NO: 1.5. The pharmaceutical composition of claim 1 , wherein the enzyme reaction is carried out at a temperature of 35° C. to 45° C. and pH of 6 to 8.6. The pharmaceutical composition of claim 1 , wherein the neoagarooligosaccharide is one or more kinds selected from the group consisting of neoagarobiose claim 1 , neoagarotetraose claim 1 , and neoagarohexaose.7. A food composition for prevention or ameliorating sepsis or septic shock claim 1 , the food composition comprising neoagarooligosaccharide prepared from agar or agarose by DagA enzyme reaction as an active ingredient.8. A pharmaceutical composition for reinforcement of immunization claim 1 , the pharmaceutical composition comprising neoagarooligosaccharide prepared from agar or agarose by a DagA enzyme reaction claim 1 , as an active ingredient.9. A functional ...

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Номер записи: 91
02-03-2017 дата публикации

Tetrameric Cytokines with Improved Biological Activity

Номер: US20170056516A1
Автор: Chang Chien-Hsing, GOLDENBERG David M., Rossi Edmund A.
Принадлежит:

The present invention concerns methods and compositions for forming cytokine-antibody complexes using dock-and-lock technology. In preferred embodiments, the cytokine-MAb DNL complex comprises an IgG antibody attached to two AD (anchor domain) moieties and four cytokines, each attached to a DDD (docking and dimerization domain) moiety. The DDD moieties form dimers that bind to the AD moieties, resulting in a 2:1 ratio of DDD to AD. The cytokine-MAb complex exhibits improved pharmacokinetics, with a significantly longer serum half-life than either naked cytokine or PEGylated cytokine. The cytokine-MAb complex also exhibits significantly improved in vitro and in vivo efficacy compared to cytokine alone, antibody alone, unconjugated cytokine plus antibody or cytokine-MAb DNL complexes incorporating an irrelevant antibody. In more preferred embodiment the cytokine is G-CSF, erythropoietin or INF-α2b. 1. A cytokine-antibody complex comprising;a) a cytokine moiety attached to a DDD (dimerization and docking domain) moiety, wherein the amino acid sequence of said DDD moiety is residues 1-44 of human protein kinase A (PKA) RIIα, wherein the cytokine moiety is selected from the group consisting of MIF (macrophage migration inhibitory factor), IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, IL-16, IL-17, IL-18, interferon-α, interferon-β, G-CSF, GM-CSF, Flt-3, erythropoietin, thrombopoietin, VEGF, FGF, insulin, and hGH; andb) an antibody moiety attached to an AD (anchor domain) moiety, wherein the amino acid sequence of the AD moiety is from a human AKAP (A-kinase anchoring protein), wherein the DDD moieties form a dimer that binds to the AD moiety to form the complex, wherein the antibody moiety binds to a human antigen selected from the group consisting of carbonic anhydrase IX, CSAp, CD1, CD1a, CD2, CD3, CD5, CD8, CD14, CD15, CD16, CD19, IGF-1R, CD20, CD21, CD22, CD23, CD25, CD30, CD33, CD37, CD38, CD40, CD45, CD46, CD52, CD55, CD59 ...

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Номер записи: 92
20-02-2020 дата публикации

METHOD AND KITS FOR IDENTIFYING OF CDK9 INHIBITORS FOR THE TREATMENT OF CANCER

Номер: US20200054640A1
Автор: Huang Chun-Hao, LOWE Scott William, LUJAMBIO Amaia, SHERR Charles J.
Принадлежит:

A method of determining sensitivity to cancer treatment includes the step of determining the presence of overexpression of MYC in a biological sample from a patient suffering from cancer, wherein the presence of overexpression of MYC indicates a sensitivity to a treatment by a CDK9 inhibitor and wherein the cancer is selected from the group consisting of carcinoma, leukemia, and lymphoma. 1. A method of determining sensitivity to cancer treatment in a patient suffering from cancer , the method comprising the steps of:determining the presence of overexpression of MYC in a biological sample from the patient, wherein the presence of overexpression of MYC indicates a sensitivity to a treatment by a CDK9 inhibitor,wherein the cancer is selected from the group consisting of carcinoma, leukemia, and lymphoma.2. The method of claim 1 , wherein the cancer is hepatocellular carcinoma.3. The method of claim 1 , wherein the cancer is non-small cell lung carcinoma.4. The method of claim 1 , further comprising the step of administering an effective amount of the CDK9 inhibitor into the patient if a overexpression of MYC is found in the biological sample.5. The method of claim 4 , wherein the CDK9 inhibitor is PHA 767491 claim 4 , PHA-793887 claim 4 , PHA-848125 claim 4 , BAY 1143572 claim 4 , BAY 1112054 claim 4 , Cdk9 inhibitor II (CAS 140651-18-9 from Calbiochem) claim 4 , DRB claim 4 , AZD-5438 claim 4 , SNS-032 claim 4 , dinaciclib claim 4 , LY2857785 claim 4 , flavopiridol claim 4 , purvalanol B claim 4 , CDKI-71 claim 4 , CDKI-73 claim 4 , CAN508 claim 4 , FIT-039 claim 4 , CYC065 claim 4 , 3 claim 4 ,4-dimethyl-5-[2-(4-piperazin-1-yl-phenylamino)-pyrimidin-4-yl]-3H-thiazol-2-one claim 4 , wogonin claim 4 , apigenin claim 4 , chrysin claim 4 , luteolin claim 4 , 4-methyl-5-[2-(3-nitroanilino)pyrimidin-4-yl]-1 claim 4 ,3-thiazol-2-amine claim 4 , shRNAs against CDK9 claim 4 , anti-sense mRNA against CDK9 and anti-CDK9 antibodies.6. A method of evaluating the efficacy of ...

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Номер записи: 93
03-03-2016 дата публикации

GLUCOSE AND XYLOSE CO-FERMENTING MICROORGANISM THAT EXPRESSES ACTIVE GLUCOAMYLASE

Номер: US20160060659A1
Автор: Ho Nancy W.Y.
Принадлежит:

Provided are microorganisms, e.g., the yeast, that have been made able to co-ferment xylose sugar-obtained from hydrolyzing plant cellulosic biomass form trees, grasses, straws, etc., with glucose that can be obtained from hydrolyzing either edible feedstocks such as starch, cane sugar, etc. or from hydrolyzing cellulose from various types of non-edible cellulosic biomass. The microorganisms are also capable of expressing an amylase, e.g., glucoamylase, having nonnegligible enzymatic activity, capable of producing glucose from oligo- or polysaccharides obtained by treating soluble starch with α-amylase. In some embodiments, nucleotidic material is provided comprising genes actively expressing xylose reductase, xylitol dehydrogenase and xylulokinase as well as an active gene expressing glucoamylase. Vectors and other compositions of matter are provided as. 1. In a container containing biomass , a microorganism capable of fermenting glucosidic and xylosic material to ethanol and also capable of expressing and/or secreting glucoamylase having nonnegligible enzymatic activity for breaking down polysaccharides of the biomass , the polysaccharides containing at least two six-carbon saccharidic unit.2. The microorganism of claim 1 , wherein the microorganism has nucleotidic material traceable to a man-made recombinant process.3. The microorganism of claim 2 , wherein the microorganism is a yeast.4. The microorganism of claim 3 , wherein the yeast ferments glucose to ethanol.5Saccharomyces.. The microorganism of claim 3 , wherein the yeast is of the genus6. The microorganism of claim 3 , wherein the microorganism is produced by a process comprising:(a) transforming yeast cells with a replicative and integrative plasmid comprising an autonomous replicating sequence, exogenous nucleotidic material, and a selection marker; and(b) repeatedly replicating the cells from step (a) to produce a number of generations of progeny cells while selecting for cells which include the ...

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Номер записи: 94
02-03-2017 дата публикации

ORGANIC ACID SYNTHESIS FROM C1 SUBSTRATES

Номер: US20170058280A1
Автор: GUARNIERI Michael T., HENARD Calvin A.
Принадлежит:

Presented herein are biocatalysts and methods for converting C1-containing materials to organic acids such as muconic acid or adipic acid. 1. An engineered cell , comprising at least one exogenously added gene encoding a 3-dehydroshikimate dehydratase , a protocatechuic acid decarboxylase , a catechol 1 ,2-dioxygenase , aroG , trpE , a phosphoenolpyruvate synthase , or a transketolase; wherein the cell is able to convert a C1 substrate to an organic acid.2. The engineered cell of claim 1 , wherein the cell comprises genes encoding a 3-dehydroshikimate dehydratase claim 1 , a protocatechuic acid decarboxylase claim 1 , and a catechol 1 claim 1 ,2-dioxygenase.3Klebsiella variicola.. The engineered cell of claim 2 , wherein the 3-dehydroshikimate dehydratase is AroZ from4Enterobacter cloacae.. The engineered cell of claim 2 , wherein the protocatechuic acid decarboxylase is AroY from5Acinetobacter.. The engineered cell of claim 2 , wherein the catechol 1 claim 2 ,2-dioxygenase is CatA from6Klebsiella variicola,Enterobacter cloacae,Acinetobacter.. The engineered cell of claim 2 , wherein the 3-dehydroshikimate dehydratase is AroZ from the protocatechuic acid decarboxylase is AroY from and the catechol 1 claim 2 ,2-dioxygenase is CatA from7. The engineered cell of claim 1 , wherein the cell further comprises an exogenously added gene encoding a phosphoketolase.8. The engineered cell of claim 7 , wherein the phosphoketolase is PktA or PktB.9. The engineered cell of claim 1 , wherein the cell further comprises an exogenously added gene encoding a CbbL claim 1 , CbbS claim 1 , CbbQ or CbbP polypeptide.10. The engineered cell of claim 9 , wherein the cell further comprises exogenously added genes encoding CbbL claim 9 , CbbS claim 9 , CbbQ and CbbP polypeptides.11. The engineered cell of claim 1 , wherein the cell is a C1 metabolizing cell.12Methylomonas, Methylobacter, Methylococcus, Methylosinus, Methylocystis, Methylomicrobium, Methanomonas, Methylophilus, Methylobacillus ...

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Номер записи: 95
04-03-2021 дата публикации

ENGINEERING CRASSULACEAN ACID METABOLISM (CAM) PATHWAYS IN PLANTS

Номер: US20210062214A1
Автор: Cushman John C., Lim Sung Don, Yim Won Cheol
Принадлежит:

Disclosed herein are methods of altering CAM pathways in plants. In some examples, a disclosed method includes overexpressing one or more genes encoding one or more enzymes that carry out the basic biochemical sequence of nocturnal COfixation (carboxylation) into Cacids (malate), store Cacids in the vacuole of the plant, and/or then decarboxylate and refix the released COby Cphotosynthesis during the subsequent day in a plant cell, thereby altering CAM in the plant cell. Also disclosed herein are isolated polynucleotide sequences, transformation vectors, transgenic plant cells, plant part, and plants. The disclosed methods and compositions can be used to improve the water-use efficiency and drought tolerance and durability of plants, such as in plants in arid environments, and also enhance the ability of plants to perform net COfixation resulting in increased biomass production and accumulation. 1Mesembryanthemum crystallinumMesembryanthemum crystallinumMesembryanthemum crystallinumMesembryanthemum crystallinum. A method of enhancing Crassulacean acid metabolism (CAM) pathways , comprising increasing expression of at least one gene encoding McBca2 (Beta-carbonic anhydrase) , at least one gene encoding McPpc1 (phosphoenolpyruvate carboxylase) , at least one gene encoding McPpck (phosphoenolpyruvate carboxylase kinase) , and at least one gene encoding McNAD-Mdh2 (NAD(P) malate dehydrogenase 2) in a plant cell as compared to expression in a control plant , thereby enhancing the carboxylation module of CAM in the plant cell.2. The method of claim 1 , wherein the at least one gene encoding McBca2 has the nucleotide sequence of SEQ ID NO: 1 claim 1 , the at least one gene encoding McPpc1 has the nucleotide sequence of SEQ ID NO: 3 claim 1 , the at least one gene encoding McPpck has the nucleotide sequence of SEQ ID NO: 2 claim 1 , and the at least one gene encoding McNAD-Mdh2 has the nucleotide sequence of SEQ ID NO: 4.3. The method of claim 1 , further comprising: ...

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Номер записи: 96
04-03-2021 дата публикации

MICROBIAL FERMENTATION FOR THE PRODUCTION OF TERPENES

Номер: US20210062229A1
Автор: Koepke Michael
Принадлежит:

The invention provides a method for producing a terpene or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces a terpene or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and/or farnesene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, or terpene biosynthesis pathways. 1ClostridiumMoorella.. A recombinant C1-fixing bacteria capable of producing mevalonic acid , or a terpene precursor , from a carbon source comprising a nucleic acid encoding a group of exogenous enzymes comprising thiolase , 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase , and HMG-CoA reductase , wherein the bacteria are or2. The bacteria according to claim 1 , further comprising a nucleic acid encoding a group of enzymes comprising mevalonate kinase claim 1 , phosphomevalonate kinase claim 1 , and mevalonate diphosphate decarboxylase.3. The bacteria according to claim 2 , wherein the terpene precursor is isopentenyl diphosphate.4. The bacteria according to claim 2 , further comprising a nucleic acid encoding an exogenous enzyme selected from the group consisting of isopentenyl diphosphate isomerase and geranyltranstransferase.5. The bacteria according to claim 2 , further comprising a nucleic acid encoding both exogenous enzymes isopentenyl diphosphate isomerase and geranyltranstransferase.6. The bacteria according to claim 4 , wherein the terpene precursor is dimethylallyl pyrophosphate or geranyl pyrophosphate.7. The bacteria according to claim 4 , further comprising a nucleic acid encoding an exogenous enzyme comprising isoprene synthase.8. The bacteria according to claim 4 , wherein the terpene precursor is farnesyl pyrophosphate.9. The bacteria according to claim 4 , further comprising a nucleic acid ...

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Номер записи: 97
05-03-2015 дата публикации

RECOMBINANT BACTERIA COMPRISING NOVEL SUCROSE TRANSPORTERS

Номер: US20150064755A1
Автор: RUEBLING-JASS KRISTIN, Tomb Jean-Francois, Van Dyk Tina K, You Zheng
Принадлежит: E I DU PONT DE NEMOURS AND COMPANY

Recombinant bacteria capable of metabolizing sucrose are described. The recombinant bacteria comprise in their genome or on at least one recombinant construct, a novel nucleotide sequence encoding a polypeptide having sucrose transporter activity and a nucleotide sequence encoding a polypeptide having sucrose hydrolase activity. These nucleotide sequences are each operably linked to the same or a different promoter. Recombinant bacteria capable of metabolizing sucrose to produce glycerol and/or glycerol-derived products such as 1,3-propanediol and 3-hydroxypropionic acid are also described. 2. The bacterium of claim 1 , wherein the polypeptide having sucrose hydrolase activity is classified as EC 3.2.1.26 or EC 2.4.1.7.3. The recombinant bacterium of further comprising in its genome or on at least one recombinant construct claim 1 , a nucleotide sequence encoding a polypeptide having fructokinase activity.4. The bacterium of claim 3 , wherein the polypeptide having fructokinase activity is classified as EC 2.7.1.4 claim 3 , EC 2.7.1.3 claim 3 , or EC 2.7.1.1.5Escherichia, Klebsiella, CitrobacterAerobacter.. The recombinant bacterium of wherein said bacterium is selected from the group consisting of the genera: claim 1 , and6Escherichia coli.. The recombinant bacterium of wherein said bacterium is7. The recombinant bacterium of wherein the recombinant bacterium produces 1 claim 1 ,3-propanediol claim 1 , glycerol claim 1 , and/or 3-hydroxypropionic acid.8. A process for making glycerol claim 1 , 1 claim 1 ,3-propanediol and/or 3-hydroxypropionic acid from sucrose comprising:{'claim-ref': {'@idref': 'CLM-00007', 'claim 7'}, 'a) culturing the recombinant bacterium of in the presence of sucrose; and'}b) optionally, recovering the glycerol, 1,3-propanediol and/or 3-hydroxypropionic acid produced. The invention relates to the fields of microbiology and molecular biology. More specifically, recombinant bacteria comprising novel sucrose transporters and methods of utilizing ...

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Номер записи: 98
09-03-2017 дата публикации

Gene Products of Bacillus Licheniformis Which Form Odorous Substances and Improved Biotechnological Production Methods Based Thereon

Номер: US20170067064A1
Автор: Bessler Cornelius, Ehrenreich Armin, Evers Stefan, Feesche Jorg, Gottschalk Gerhard, Henne Anke, Herzberg Christina, Liesegang Heiko, Maurer Karl-Heinz, Veith Birgit
Принадлежит:

The present invention relates to 25 hitherto undescribed genes of and gene products derived thereform and all sufficiently homologous nucleic acids and proteins thereof. They occur in five different metabolic pathways for the formation of odorous substances. The metabolic pathways in question are for the synthesis of: 1) isovalerian acid (as part of the catabolism of leucine), 2) 2-methylbutyric acid and/or isobutyric acid (as part of the catabolism of valine and/or isoleucine), 3) butanol and/or butyric acid (as part of the metabolism of butyric acid), 4) propyl acid (as part of the metabolism of propionate) and/or 5) cadaverine and/or putrescine (as parts of the catabolism of lysine and/or arginine). The identification of these genes allows biotechnological production methods to be developed that are improved to the extent that, to assist these nucleic acids, the formation of the odorous substances synthesised via these metabolic pathways can be reduced by deactivating the corresponding genes in the micro-organism used for the biotechnological production. In addition, these gene products are thus available for preparing reactions or for methods according to their respective biochemical properties. 1. A nucleic acid selected from the group consisting of:(a) nucleic acids coding for a gene product (putative branched-chain amino acid aminotransferase; E.C. 2.6.1.42) involved in the synthesis of 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 67% identity to the nucleotide sequence indicated in SEQ ID NO. 1;(b) nucleic acids coding for a gene product (putative branched-chain amino acid aminotransferase; E.C. 2.6.1.42) involved in the synthesis of 2-methylbutyric acid and/or isobutyric acid and having a nucleotide sequence which shows at least 78% identity to the nucleotide sequence indicated in SEQ ID NO. 3;(c) nucleic acids speA coding for a gene product (lysine and/or arginine decarboxylase; E.C. 4.1.1.18 or 4.1.1.19 ...

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Номер записи: 99
11-03-2021 дата публикации

FERMENTATION PROCESS FOR IMPROVED GLYCEROL AND ACETIC ACID CONVERSION

Номер: US20210071205A1
Автор: DE BRUIJN Hans Marinus Charles Johannes, de Laat Wilhelmus Theodorus Antonius Maria, Klaassen Paul
Принадлежит:

The invention relates to a process for producing a fermentation product that comprises fermentation of a carbon source in a reactor with a cell, capable of converting sugar, glycerol and acetic acid, wherein the carbon source comprises sugar and acetic acid, comprising the following steps: 1. Process for producing a fermentation product that comprises fermentation of a carbon source in a reactor with a cell , capable of converting sugar , glycerol and acetic acid , wherein the carbon source comprises sugar and acetic acid , comprising:a) Inoculating a optionally diluted carbon source with the cell;b) optionally fermenting the reactor in batch mode;c) adding carbon source comprising glycerol and optionally sugar gradually to the reactor;d) after sufficient fermentation time, isolation of fermentation product from the reactor,e) optionally keeping the remaining fraction after isolation of d) as spent broth; andf) optionally using the spent broth in a) to dilute the carbon source.2. Process according to claim 1 , wherein the carbon source comprises lignocellulosic hydrolysate.3. Process according to claim 2 , wherein the amount of glycerol added is such that the molar concentration of glycerol in the reactor is about twice the molar concentration or 1.8 to 2.2 times the molar concentration of acetic acid in the reactor.4. Process according to claim 2 , wherein the added glycerol originates from a starch or sugar based ethanol product plant or a biodiesel plant.5. Process according to claim 1 , wherein the addition of glycerol is commenced when the glucose concentration in reactor is 2 g/l or lower.6. Process according to claim 1 , wherein the remaining fraction after isolation of f) is kept as spent broth; and the spent broth is used in b).7. Process according claim 1 , wherein the cell is capable of consuming xylose in the lignocelluloic hydrolysate claim 1 , optionally substantially all xylose.8. Process according to claim 1 , wherein the cell is a yeast cell that is ...

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Номер записи: 100