MULTILAMELLAR LIPID VESICLE COMPOSITIONS INCLUDING A CONJUGATED ANAPLASTIC LYMPHOMA KINASE (ALK) VARIANT AND USES THEREOF

The invention provides compositions including stabilized multilamellar lipid vesicles having crosslinked lipid bilayers (referred to herein as interbilayer-crosslinked multilamellar vesicles or ICMV) and including an ALK variant, pharmaceutical compositions containing vesicles (e.g., ICMV) including an ALK variant, and methods of treatment using such compositions. The invention provides compositions including stabilized multilamellar lipid vesicles with crosslinked lipid bilayers (e.g., an interbilayer-crosslinked multilamellar vesicle or ICMV) containing an Anaplastic lymphoma kinase (ALK) variant as an antigen that is associated with solid tumor cancers. 1. A composition comprising:(a) a multilamellar lipid vesicle having crosslinks between lipid bilayers; and(b) an anaplastic lymphoma kinase (ALK) variant.2. The composition of claim 1 , wherein said ALK variant is conjugated to a lipid.3. The composition of or claim 1 , wherein said composition further comprises a nucleophosmin (NPM) protein or a fragment thereof.4. The composition of claim 3 , wherein said fragment of said NPM protein is an extracellular domain of said NPM protein.5. The composition of or claim 3 , wherein said NPM protein is fused to said ALK variant.6. The composition of or claim 3 , wherein said composition further comprises a tropomyosin (TMP3) protein or a fragment thereof.7. The composition of claim 6 , wherein said fragment of said TMP3 protein is an extracellular domain of said TMP3 protein.8. The composition of or claim 6 , wherein said TMP3 protein is fused to said ALK variant.9. The composition of or claim 6 , wherein said composition further comprises a 5-Aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC) protein or a fragment thereof.10. The composition of claim 9 , wherein said fragment of said ATIC protein is an extracellular domain of said ATIC protein.11. The composition of or claim 9 , wherein said ATIC protein is fused to said ALK variant. ...

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

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 ...

GENETICALLY MODIFIED MICROORGANISMS HAVING IMPROVED TOLERANCE TOWARDS L-SERINE

The present invention generally relates to the microbiological industry, and specifically to the production of L-serine or L-serine derivatives using genetically modified bacteria. The present invention provides genetically modified microorganisms, such as bacteria, wherein the expression of genes encoding for enzymes involved in the degradation of L-serine is attenuated, such as by inactivation, which makes them particularly suitable for the production of L-serine at higher yield. The present invention also provides means by which the microorganism, and more particularly a bacterium, can be made tolerant towards higher concentrations of serine. The present invention also provides methods for the production of L-serine or L-serine derivative using such genetically modified microorganisms. 1. A bacterium , which has been modified to attenuate expression of at least one gene coding for a polypeptide comprising serine deaminase activity and/or to attenuate expression of a gene coding for a polypeptide comprising serine hydroxymethyltransferase activity.235-. (canceled)36. The bacterium according to claim 1 , wherein the at least one gene coding for a polypeptide comprising serine deaminase activity is selected from the group consisting of sdaA claim 1 , sdaB and tdcG.37. The bacterium according to claim 1 , wherein the gene coding for a polypeptide comprising serine hydroxymethyltransferase activity is glyA.38. The bacterium according to claim 1 , wherein the bacterium has been modified to attenuate expression of the genes sdaA claim 1 , sdaB claim 1 , tdcG and glyA.39. The bacterium according to claim 1 , wherein the bacterium has been modified to attenuate expression of at most three of the genes sdaA claim 1 , sdaB claim 1 , tdcG and glyA.40. The bacterium according to claim 1 , wherein the expression of the gene or genes is attenuated by inactivation of the gene or genes.41. The bacterium according to claim 1 , wherein at least one gene coding for a polypeptide ...

VITAMIN PROTOTROPHY AS A SELECTABLE MARKER

One or more genes in a biosynthesis pathway for a vitamin or other essential nutrient which is needed for the survival of a microorganism can be used as an effective selective marker to identify cells transformed with an exogenous nucleic acid. The microorganism does not naturally contain or express the one or more gene. This permits genetic manipulations to be performed. It permits lower cost fermentations to be performed. It permits production of the essential nutrient for subsequent commodity use. 1. A method of using vitamin prototrophy as a selectable marker comprising:(a) transforming a microorganism auxotrophic for a vitamin with both a vitamin biosynthesis gene and a gene conferring a desired property, wherein the microorganism is not transformed with an antibiotic resistance gene and(b) culturing the microorganism in the absence of the vitamin,wherein growth of the microorganism in the absence of the vitamin indicates successful transformation of the microorganism with both the vitamin biosynthesis gene and the gene conferring a desired property.2. The method of claim 1 , wherein the culturing is performed in the absence of an antibiotic.3. The method of claim 1 , wherein antibiotic resistance is not used as a selectable marker.4. The method of claim 1 , wherein transformation with the vitamin biosynthesis gene renders the microorganism prototrophic for the vitamin.5. The method of claim 1 , wherein the desired property is not selectable.6. The method of claim 1 , wherein the vitamin is thiamine.7. The method of claim 1 , wherein the vitamin biosynthesis gene encodes thiamine biosynthesis protein (ThiC) (EC 4.1.99.17).8Clostridium ragsdalei.. The method of claim 1 , wherein the vitamin biosynthesis gene is thiC derived from9. The method of claim 1 , wherein the vitamin is pantothenate.10. The method of claim 1 , wherein the vitamin biosynthesis gene encodes methyl-2-oxobutanoate hydroxymethyltransferase (PanB) (EC 2.1.2.11) claim 1 , pantoate-beta-alanine ...

METHOD FOR THE PRODUCTION OF L-SERINE USING GENETICALLY ENGINEERED MICROORGANISMS DEFICIENT IN SERINE DEGRADATION PATHWAYS

The present invention generally relates to the microbiological industry, and specifically to the production of L-serine using genetically modified bacteria. The present invention provides genetically modified microorganisms, such as bacteria, wherein the expression of genes encoding for enzymes involved in the degradation of L-serine is attenuated, such as by inactivation, which makes them particularly suitable for the production of L-serine at higher yield. The present invention also provides means by which the microorganism, and more particularly a bacterium, can be made tolerant towards higher concentrations of serine. The present invention also provides methods for the production of L-serine or L-serine derivative using such genetically modified microorganisms. 1. A bacterium , which has been modified to attenuate expression of genes coding for polypeptides comprising serine deaminase activity and to attenuate expression of a gene coding for a polypeptide comprising serine hydroxymethyltransferase activity.220-. (canceled)21. The bacterium according to claim 1 , wherein the expression of the genes sdaA claim 1 , sdaB claim 1 , tdcG and glyA is attenuated.22. The bacterium according to claim 1 , wherein the expression of the genes is attenuated by inactivation of the genes.23. The bacterium according to claim 1 , wherein said bacterium has been further modified to overexpress a 3-phosphoglycerate dehydrogenase claim 1 , a phosphoserine phosphatase and a phosphoserine aminotransferase.24. The bacterium according to claim 1 , wherein said bacterium is capable of growing in a minimal culture medium comprising L-serine at a concentration of at least about 6.25 g/L.25. The bacterium according to claim 1 , wherein said bacterium is capable of growing in a minimal culture medium comprising L-serine at a concentration of at least about 6.25 g/L at a growth rate of at least 0.1 hrduring exponential growth.26. The bacterium according to claim 1 , wherein said bacterium ...

Stable gene transfer to proliferating cells

Provided herein are methods for facilitating or inducing stable transgene integration and expression in a proliferating cell, comprising administering to the cell (i) a recombinant AAV (rAAV) vector comprising the transgene flanked by transposon-derived inverted terminal repeat sequences, which sequences are in turn flanked by AAV-derived inverted terminal repeat regions, and (ii) a source of a transposase that recognises said transposon-derived inverted terminal repeat sequences and directs the genomic integration of the transgene into the genome of the proliferating cell. Also provide are methods and transgene delivery systems for the treatment or prevention of diseases affecting, associated with or characterised by proliferating cells.

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Non-viral delivery systems comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A composition comprising a non-viral carrier and a nucleic acid molecule comprising an engineered sequence encoding human ornithine transcarbamylase (hOTCase) and expression control sequences which direct expression of hOTCase in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over at least the mature hOTC sequence of SEQ ID NO: 1 , wherein said hOTC nucleic acid sequence is selected from the nucleic acid sequence comprising SEQ ID NO: 5 or a nucleic acid sequence at least about 96 to about 99% identical thereto , a nucleic acid sequence selected from SEQ ID NO: 9 , or a nucleic acid sequence at least about 96 to about 99% identical thereto , a nucleic acid sequence comprising SEQ ID NO: 11 or a nucleic acid sequence at least about 96 to about 99% identical thereto , or a nucleic acid sequence comprising SEQ ID NO: 13 or a nucleic acid sequence at least about 96 to about 99% identical thereto.2. The composition according to claim 1 , wherein the hOTC nucleic acid sequence is a DNA sequence of SEQ ID NO: 4.3. The composition according to claim 1 , wherein the hOTC nucleic acid sequence is a DNA sequence of SEQ ID NO: 3.4. The composition according to claim 1 , wherein the hOTC nucleic acid sequence is a DNA sequence of SEQ ID NO: 8.5. The composition according to claim 1 , wherein the hOTC is an RNA sequence of SEQ ID NO: 10.6. The composition according to claim 1 , wherein the hOTC is an RNA sequence of SEQ ID NO: 12.7. The composition according to claim 1 , wherein the hOTC is a chimeric OTC comprises a ...

mRNA FOR USE IN TREATMENT OF HUMAN GENETIC DISEASES

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification 5 conferring stability to the RNA, as well as related methods, are disclosed. 1. A composition for modulating the expression of a protein in a target cell , wherein said composition comprises at least one RNA molecule and a transfer vehicle and wherein the RNA comprises at least one modification which confers stability to the RNA.2. The composition of claim 1 , wherein the RNA molecule is selected from the group consisting of mRNA claim 1 , miRNA claim 1 , snRNA claim 1 , and snoRNA.3. The composition of claim 1 , wherein the RNA molecule comprises more than one modification which confers stability to the RNA molecule.4. The composition of claim 1 , wherein the RNA molecule comprises a modification of the 5′ untranslated region of said RNA molecule.5. The composition of claim 4 , wherein said modification comprises a partial sequence of a CMV immediate-early 1 (IE 1) gene.6. The composition of claim 5 , wherein said partial sequence of the CMV immediate-early 1 (IEI) gene comprises SEQ ID NO: 2 or SEQ ID NO: 1.7. The composition of claim 4 , wherein said modification comprises the inclusion of a poly A tail.8. The composition of claim 4 , wherein said modification comprises the inclusion of a Cap 1 structure.9. The composition of claim 1 , wherein the RNA molecule comprises a modification of the 3′ untranslated of said RNA molecule.10. The composition of claim 9 , wherein said modification comprises the inclusion of a sequence encoding human growth hormone (hGH).11. The composition of claim 10 , wherein said sequence encoding human growth hormone (hGH) comprises SEQ ID NO: 3.12. The composition of claim 9 , wherein said modification comprises the inclusion of a poly A tail.13. The composition of claim 1 , wherein the RNA encodes ornithine carbamoyl transferase.14. The composition of claim 1 , wherein the RNA encodes ...

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

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

MESSENGER UNA MOLECULES AND USES THEREOF

This invention provides a range of translatable messenger UNA (mUNA) molecules. The mUNA molecules can be translated in vitro and in vivo to provide an active polypeptide or protein, or to provide an immunization agent or vaccine component. The mUNA molecules can be used as an active agent to express an active polypeptide or protein in cells or subjects. Among other things, the mUNA molecules are useful in methods for treating rare diseases. 1. A mUNA molecule , comprising one or more UNA monomers , and comprising nucleic acid monomers , wherein the mUNA molecule is translatable to express a polypeptide or protein.2. The molecule of claim 1 , wherein the molecule comprises from 200 to 12 claim 1 ,000 monomers.3. The molecule of claim 1 , wherein the molecule comprises from 200 to 4 claim 1 ,000 monomers.4. The molecule of claim 1 , wherein the molecule comprises from 1 to 8 claim 1 ,000 UNA monomers.5. The molecule of claim 1 , wherein the molecule comprises from 1 to 100 UNA monomers.6. The molecule of claim 1 , wherein the molecule comprises from 1 to 20 UNA monomers.7. The molecule of claim 1 , wherein the molecule comprises one or more modified nucleic acid nucleotides claim 1 , or one or more chemically-modified nucleic acid nucleotides.8. The molecule of claim 1 , wherein the molecule comprises a 5′ cap claim 1 , a 5′ untranslated region of monomers claim 1 , a coding region of monomers claim 1 , a 3′ untranslated region of monomers claim 1 , and a tail region of monomers.9. The molecule of claim 8 , wherein the molecule comprises a translation enhancer in a 5′ or 3′ untranslated region.10. The molecule of claim 1 , wherein the molecule is translatable in vivo.11. The molecule of claim 1 , wherein the molecule is translatable in vitro.12. The molecule of claim 1 , wherein the molecule is translatable in a mammalian cell.13. The molecule of claim 1 , wherein the molecule is translatable in a human in vivo.14. The molecule of claim 1 , wherein a translation ...

Compositions useful in treatment of ornithine transcarbamylase (otc) deficiency

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis cirrhosis in OTCD patients by administering hOTC.

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

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 ...

MULTIMERIC CODING NUCLEIC ACID AND USES THEREOF

The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends. 1. A multimeric coding nucleic acid (MCNA) comprising two polynucleotides linked via 3′ ends via an oligonucleotide bridge comprising a stable 3-3′ inverted phosphodiester linkage such that the multimeric coding nucleic acid compound comprises two or more 5′ ends , andwherein at least one of the two polynucleotides is an encoding polynucleotide.24.-. (canceled)5. The MCNA of claim 1 , wherein the at least one encoding polynucleotide encodes a protein of interest.6. The MCNA of claim 5 , wherein the two polynucleotides are encoding polynucleotides claim 5 , and each of the two encoding polynucleotides encodes the same protein.7. The MCNA of claim 5 , wherein the two polynucleotides are encoding polynucleotides claim 5 , and each of the two encoding polynucleotides encodes a distinct protein.811.-. (canceled)12. The MCNA of claim 1 , wherein the at least one encoding polynucleotide comprises a 3′ UTR.13. (canceled)14. The MCNA of claim 12 , wherein the 3′ UTR comprises a plurality of multi-A segments with spacers in between.1526.-. (canceled)27. The MCNA of claim 1 , wherein the nucleosides comprising the oligonucleotide bridge are selected from the group consisting of 2′-OMe-A claim 1 , 2′-OMe-G claim 1 , 2′-OMe-C claim 1 , 2′-OMe-U claim 1 , 2′-F-A claim 1 , 2′-F-G claim 1 , 2′-F-C claim 1 , 2′-F-U claim 1 , LNA-A claim 1 , LNA-G claim 1 , LNA-C claim 1 , LNA-U claim 1 , N6-methyl-adenosine claim 1 , 2-thiouridine (2sU) claim 1 , 5-methyl-cytidine (5mC) claim 1 , pseudouridine (ψU) claim 1 , and 1-methyl-pseudouridine.2830.-. (canceled)31. The MCNA of claim 1 , wherein the at least one encoding polynucleotide ...

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A recombinant viral vector comprising a nucleic acid sequence encoding human ornithine transcarbamylase (hOTC) and expression control sequences which direct expression of hOTC in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over the mature sequence or full length hOTC of SEQ ID NO: 1 , and expresses a functional hOTC , wherein said hOTC nucleic acid sequence is SEQ ID NO: 9 or a nucleic acid sequence at least 96 to 99% identical thereto.2. A recombinant viral vector comprising a nucleic acid sequence encoding hOTC and expression control sequences which direct expression of hOTC in a liver cell , wherein the hOTC nucleic acid sequence expresses a functional hOTC , wherein said hOTC nucleic acid sequence is SEQ ID NO: 3. This is a continuation of U.S. patent application Ser. No. 15/122,853, filed Aug. 31, 2016, which is a national stage application under 35 U.S.C. 371 of PCT/US2015/019513, filed on Mar. 9, 2015, now expired, which claims the benefit of U.S. Patent Application No. 61/950,157, filed Mar. 9, 2014, now expired. These applications are incorporated by reference in their entirety.This invention was made with support under grant Nos. P01-HD057247, P01-HL059407, and P30-DK047757 awarded by the National Institutes of Health. The US government has certain rights in the invention.Ornithine transcarbamylase (OTC) deficiency accounts for nearly half of all cases of inborn errors of urea synthesis, with a prevalence estimated to be at least 1 in 15,000. Urea cycle defects put patients at risk of life threatening ...

Secretagogues derived from oxalobacter formigenes

The present invention relates to a secretagogue compound derived from oxalate degrading bacteria, for use in the treatment of an oxalate related disease and/or oxalate related imbalance in a subject, wherein the administration of the secretagogue results in a reduction of urinary oxalate and/or plasma oxalate in the subject. The invention further relates to a pharmaceutical composition comprising such a secretagogue compound, a method for treating a subject suffering from an oxalate related disease, and to a method for preparing a secretagogue.

SECRETAGOGUES DERIVED FROM OXALOBACTER FORMIGENES

The present invention relates to a secretagogue compound derived from oxalate degrading bacteria, for use in the treatment of an oxalate related disease and/or oxalate related imbalance in a subject, wherein the administration of the secretagogue results in a reduction of urinary oxalate and/or plasma oxalate in the subject. The invention further relates to a pharmaceutical composition comprising such a secretagogue compound, a method for treating a subject suffering from an oxalate related disease, and to a method for preparing a secretagogue. 113-. (canceled)14. A pharmaceutical product comprising:{'i': 'Oxalobacter formigenes', '(i) an isolated secretagogue derived from , and'}{'i': 'Oxalobacter formigenes', '(ii) oxalate-degrading bacteria.'}15Oxalobacter formigenes. A pharmaceutical product according to claim 14 , wherein the isolated secretagogue and the oxalate-degrading bacteria are provided in separate compositions.16. The pharmaceutical product according to claim 14 , further comprising one or more selected from oxalate-degrading enzymes claim 14 , enzymes involved in oxalate metabolism claim 14 , cofactors selected from vitamin B claim 14 , NAD claim 14 , NADP claim 14 , FAD claim 14 , CoA claim 14 , ATP and ADP claim 14 , substrates selected from oxalate claim 14 , glyoxylate claim 14 , and 4-hydroxy-2-oxoglutarate claim 14 , and combinations of any thereof.17. The pharmaceutical product according to claim 14 , wherein the isolated secretagogue is selected from recombinantly expressed secretagogues and secretagogues extracted from conditioned media.18. The pharmaceutical product of claim 14 , wherein the compositions are formulated for a route of administration selected from enteral claim 14 , parenteral and topical route.19. The pharmaceutical product according to claim 14 , wherein the amino acid sequence of the isolated secretagogue has at least 85% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1-19.20. The pharmaceutical ...

P AERUGINOSA PCRV-LINKED ANTIGEN VACCINES

The present invention discloses a conjugate comprising an antigen (for example a saccharide antigen) covalently linked to a PcrV carrier protein comprising an amino acid sequence which is at least 80% identical to the sequence of SEQ ID NO:1-4, wherein the antigen is linked (either directly or through a linker) to an amino acid residue of the PcrV carrier protein. The invention also discloses PcrV proteins that contain glycosylation site consensus sequences. 171-. (canceled)72Pseudomonas aeruginosaP. aeruginosa. A conjugate comprising or consisting of an antigen covalently linked to a PcrV carrier protein comprising an amino acid sequence which is at least 70% or 80% identical to the sequence of SEQ ID NO:1-4 , wherein the antigen is linked (either directly or through a linker) to an asparagine amino acid residue of the PcrV carrier protein wherein the asparagine residue is part of a D/E-X-N-X-S/T consensus sequence introduced into the amino acid sequence which is at least 70% or 80% identical to the sequence of SEQ ID NO:1-4 , wherein X is any amino acid apart from proline , wherein the asparagine residue is situated at a position equivalent to between amino acids 24-166 or amino acids 281-317 or at amino acid 317 of SEQ ID NO:3 or at a position equivalent to between amino acids 1-143 or amino acids 258-294 or at amino acid 294 of SEQ ID NO:4.73. The conjugate of claim 72 , wherein the asparagine residue is part of a D/E-X-N-X-S/T consensus sequence claim 72 , wherein X is any amino acid apart from proline claim 72 , wherein the asparagine residue is not introduced by mutation into the sequence of SEQ ID NO:5 or a sequence having at least 80% identity to SEQ ID NO:5.74. The conjugate of claim 72 , wherein a peptide comprising the D/E-X-N-X-S/T consensus sequence is introduced into the amino acid sequence by the removal of a PcrV peptide sequence and its replacement with the peptide comprising the D/E-X-N-X-S/T consensus sequence claim 72 , preferably wherein the ...

GENETICALLY MODIFIED MICROORGANISMS HAVING IMPROVED TOLERANCE TOWARDS L-SERINE

The present invention generally relates to the microbiological industry, and specifically to the production of L-serine or L-serine derivatives using genetically modified bacteria. The present invention provides genetically modified microorganisms, such as bacteria, wherein the expression of genes encoding for enzymes involved in the degradation of L-serine is attenuated, such as by inactivation, which makes them particularly suitable for the production of L-serine at higher yield. The present invention also provides means by which the microorganism, and more particularly a bacterium, can be made tolerant towards higher concentrations of serine. The present invention also provides methods for the production of L-serine or L-serine derivative using such genetically modified microorganisms. 1. (canceled)2Escherichia coli. An bacterium which expresses a polypeptide having an amino acid sequence , which has at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 12 , wherein in said amino acid sequence at position 143 D is replaced by another amino acid.3. The bacterium according to which expresses a polypeptide having an amino acid sequence claim 2 , which has at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 12 claim 2 , wherein in said amino acid sequence at position 143 D is replaced by G.4. The bacterium according to claim 2 , wherein said bacterium comprises within the lrp gene one of more nucleotide substitutions resulting in an amino acid substitution in the encoded polypeptide at position 143.5. The bacterium according to claim 2 , wherein at least one gene selected from sdaA claim 2 , sdaB claim 2 , tdcG or glyA is inactivated.6. The bacterium according to claim 2 , wherein at least two genes selected from sdaA claim 2 , sdaB claim 2 , tdcG or glyA are inactivated.7. The bacterium according to claim 2 , wherein at least three genes selected from sdaA claim 2 , sdaB claim 2 , tdcG or glyA are inactivated ...

Disease model pig exhibiting stable phenotype, and production method thereof

Disease model pigs produced by nuclear transplantation, disease model pigs exhibiting stable phenotypes and production methods thereof are provided. Chimeric pigs for producing disease model pigs exhibiting stable phenotypes, genital glands thereof, and germ cells thereof are also provided. A method for producing a genetically modified disease model pig, includes: (a) transplanting a nucleus of a genetically modified cell into cytoplasm of an egg; (b) developing an obtained clonal embryo in a womb of a female pig to obtain an offspring; and mating the obtained offspring or having the offspring undergo sexual reproduction to further obtain the genetically modified offspring as a disease model pig.

MICROORGANISM HAVING ENHANCED L-THREONINE PRODUCING ABILITY AND METHOD FOR PRODUCING THREONINE USING THE SAME

The present application relates to a microorganism having an enhanced L-threonine producing ability and a method for producing L-threonine using the same. 1Corynebacterium. A microorganism of the genus for producing L-threonine having an enhanced glycine transporter activity.2Corynebacterium ammoniagenes.. The microorganism of claim 1 , wherein the glycine transporter is derived from3. The microorganism of claim 1 , wherein the glycine transporter protein is CycA protein.4. The microorganism of claim 1 , wherein the protein comprises an amino acid sequence of SEQ ID NO: 16 or an amino acid sequence having at least 90% homology thereto.5. The microorganism of claim 1 , wherein the microorganism has an activity of glycine cleavage protein which is further enhanced.6. The microorganism of claim 1 , wherein the glycine cleavage protein is at least one selected from the group consisting of GcvP claim 1 , GcvT claim 1 , GcvH claim 1 , LipB claim 1 , and LipA.7Corynebacterium ammoniagenes.. The microorganism of claim 6 , wherein the glycine cleavage protein is derived from8. The microorganism of claim 6 , wherein the GcvP comprises an amino acid sequence of SEQ ID NO: 38 claim 6 , GcvT comprises an amino acid sequence of SEQ ID NO: 39 claim 6 , GcvH comprises an amino acid sequence of SEQ ID NO: 40 claim 6 , LipA comprises an amino acid sequence of SEQ ID NO: 41 claim 6 , and LipB comprises an amino acid sequence of SEQ ID NO: 42 claim 6 , or an amino acid sequence having at least 90% homology to the respective amino acid9CorynebacteriumCorynebacterium glutamicum.. The microorganism of claim 1 , wherein the microorganism of the genus for producing L-threonine is10. A composition for producing L-threonine claim 1 , comprising the microorganism of .11. A method for producing L-threonine claim 1 , comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'culturing the microorganism of ; and'}recovering L-threonine from the microorganism or medium.12Corynebacterium. Use of ...

L-ORNITHINE PRODUCTION IN EUKARYOTIC CELLS

The present invention relates to the provision of genetically modified microbial cells, such as yeast cells with an improved ability for producing L-ornithine and its derivatives. Overproduction of L-ornithine is obtained in the first place by the down-regulation or attenuation of specially selected genes, wherein said genes encode enzymes involved in the L-ornithine consumption and/or degradation pathways. Further L-ornithine production ability is improved by down-regulation, attenuation, deletion or overexpression of specially selected genes, wherein said genes encode enzymes and/or proteins involved in the L-ornithine ‘acetylated derivatives cycle’, L-glutamate synthesis pathways, subcellular trafficking, TCA cycle, pyruvate carboxylation pathway, respiratory electron-transport chain, and the carbon substrates' assimilation machinery. The invention additionally provides a method to produce L-ornithine with said modified eukaryotic cells. 139-. (canceled)40. A genetically modified eukaryotic cell modified for enhanced L-ornithine biosynthesis from α-ketoglutarate , wherein (a) a gene encoding a cytosolic glutamate N-acetyltransferase,', '(b) a gene encoding a cytosolic N-acetylglutamate kinase,', '(c) a gene encoding a cytosolic N-acetyl-gamma-glutamyl-phosphate reductase,', '(d) a gene encoding a cytosolic acetylornithine aminotransferase and', '(e) gene encoding a cytosolic ornithine acetyltransferase; or, '(1) the L-ornithine biosynthesis from α-ketoglutarate is cytosolic and the eukaryotic cell is modified to comprise at least one of the genes selected from a group consisting of'}(2) the eukaryotic cell is genetically modified for overexpression of at least one gene selected from the group consisting of:(a) a gene encoding an N-acetylglutamate kinase and N-acetyl-gamma-glutamyl-phosphate reductase;(b) a gene encoding a mitochondrial ornithine acetyltransferase;(c) a gene encoding an acetylornithine aminotransferase; and(d) a gene encoding an acetylglumate ...

Compositions for the prevention and treatment of parkinson's disease

Methods of preventing or retarding or reversing or abolishing the onset of Parkinson's and other neurodegenerative diseases are discussed.

Production of malonyl-coa derived products via anaerobic pathways

The present invention provides for novel metabolic pathways to convert biomass and other carbohydrate sources to malonyl-CoA derived products, such as hydrocarbons and other bioproducts, under anaerobic conditions and with the net production of ATP. More specifically, the invention provides for a recombinant microorganism comprising one or more native and/or heterologous enzymes that function in one or more engineered metabolic pathways to achieve conversion of a carbohydrate source to, e.g., long-chain hydrocarbons and hydrocarbon derivatives, wherein the one or more native and/or heterologous enzymes is activated, upregulated, downregulated, or deleted. The invention also provides for processes to convert biomass to malonyl-CoA derived products which comprise contacting a carbohydrate source with a recombinant microorganism of the invention.

MESSENGER UNA MOLECULES AND USES THEREOF

This invention provides a range of translatable messenger UNA (mUNA) molecules. The mUNA molecules can be translated in vitro and in vivo to provide an active polypeptide or protein, or to provide an immunization agent or vaccine component. The mUNA molecules can be used as an active agent to express an active polypeptide or protein in cells or subjects. Among other things, the mUNA molecules are useful in methods for treating rare diseases. 1. A mUNA molecule , comprising one or more UNA monomers , and comprising nucleic acid monomers , wherein the mUNA molecule is translatable to express a polypeptide or protein.2. The molecule of claim 1 , wherein the molecule comprises from 200 to 12 claim 1 ,000 monomers.3. The molecule of claim 1 , wherein the molecule comprises from 200 to 4 claim 1 ,000 monomers.4. The molecule of claim 1 , wherein the molecule comprises from 1 to 8 claim 1 ,000 UNA monomers.5. The molecule of claim 1 , wherein the molecule comprises from 1 to 100 UNA monomers.6. The molecule of claim 1 , wherein the molecule comprises from 1 to 20 UNA monomers.7. The molecule of claim 1 , wherein the molecule comprises one or more modified nucleic acid nucleotides claim 1 , or one or more chemically-modified nucleic acid nucleotides.8. The molecule of claim 1 , wherein the molecule comprises a 5′ cap claim 1 , a 5′ untranslated region of monomers claim 1 , a coding region of monomers claim 1 , a 3′ untranslated region of monomers claim 1 , and a tail region of monomers.9. The molecule of claim 8 , wherein the molecule comprises a translation enhancer in a 5′ or 3′ untranslated region.10. The molecule of claim 1 , wherein the molecule is translatable in vivo.11. The molecule of claim 1 , wherein the molecule is translatable in vitro.12. The molecule of claim 1 , wherein the molecule is translatable in a mammalian cell.13. The molecule of claim 1 , wherein the molecule is translatable in a human in vivo.14. The molecule of claim 1 , wherein a translation ...

Byosynthetic Production of Acyl Amino Acids

The present invention relates to a cell for producing acyl glycinates wherein the cell is genetically modified to comprise 115-. (canceled)16. A cell for producing acyl glycinates , wherein said cell is genetically modified to comprise:a) at least a first genetic mutation that, relative to the wild type cell, increases the expression of an amino acid-N-acyl-transferase;b) at least a second genetic mutation that, relative to the wild type cell, increases the expression of an acyl-CoA synthetase; andc) at least a third genetic mutation that, relative to the wild type cell, decreases the expression of at least one enzyme selected from the group consisting of: an enzyme of the glycine cleavage system; glycine hydroxymethyltransferase (GlyA); threonine aldolase (LtaE); threonine dehydrogenase (Tdh); 2-Amino-3-Ketobutyrate CoA-Ligase (Kbl); and allothreonine dehydrogenase (YdfG).17. The cell of claim 16 , wherein said cell comprises a mutation in an enzyme from the glycine cleavage system selected from the group consisting of: glycine cleavage system T protein; glycine cleavage system H protein; and glycine cleavage system P protein.18. The cell of claim 16 , wherein the third genetic mutation decreases the expression relative to the wild type cell of the glycine hydroxymethyltransferase (GlyA) claim 16 , the threonine aldolase (LtaE) claim 16 , the glycine cleavage system T protein claim 16 , the glycine cleavage system H protein and the glycine cleavage system P protein.19. The cell of claim 18 , wherein the glycine cleavage system T protein has 85% sequence identity to SEQ ID NO:58 claim 18 , the glycine cleavage system H protein has 85% sequence identity to SEQ ID NO:59 claim 18 , and the glycine cleavage system P protein has 85% sequence identity to SEQ ID NO:60.20. The cell of claim 16 , wherein the glycine hydroxymethyltransferase (GlyA) has 85% sequence identity to SEQ ID NO:61 and the threonine aldolase (LtaE) has 85% sequence identity to SEQ ID NO:62.21. The ...

Systems, Methods And Compositions For Recombinant In Vitro Transcription And Translation Utilizing Thermophilic Proteins

Another aim of the current invention may include a recombinant cell-free expression system, the reaction mixture containing all the cell-free reaction components necessary for the in vitro biosynthesis of biological compounds, proteins, enzymes, biosimilars or chemical modification of small molecules. 1. A system for recombinant cell-free expression comprising: a plurality of initiation factors (IFs);', 'a plurality of elongation factors (EFs);', 'a plurality of peptide release factors (RFs);', 'at least one ribosome recycling factor (RRF);', 'a plurality of aminoacyl-tRNA-synthetases (RSs); and', 'at least one methionyl-tRNA transformylase (MTF);, 'a core recombinant protein mixture having at least the following componentsat least one nucleic acid synthesis template;a reaction mixture having cell-free reaction components necessary for in vitro macromolecule synthesis; andwherein the above components are situated in a bioreactor configured for cell-free expression of macromolecules.2. The system of claim 1 , wherein the components of said core recombinant protein mixture comprises a core recombinant protein mixture derived from a bacteria.3. The system of claim 2 , wherein said core recombinant protein mixture derived from bacteria comprises a core recombinant protein mixture wherein at least one components is derived from a thermophilic bacteria.4Geobacillus. The system of any one of claim 2 , and claim 2 , wherein said thermophilic bacteria comprises a thermophilic Bacillaceae bacteria claim 2 , or thermophilic bacteria.5GeobacillusGeobacillus subterraneus,Geobacillus stearothermophilus.. The system of claim 4 , wherein said thermophilic bacteria is selected from the group consisting of: and6. The system of claim 1 , wherein said core recombinant protein mixture derived from bacteria comprises a core recombinant protein mixture wherein at least one components is derived from a non-thermophilic bacteria claim 1 , or a combination of non-thermophilic and ...

ADENO-ASSOCIATED VIRUS (AAV) SEROTYPE 8 SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. 1. An adeno-associated virus (AAV)8 viral vector comprising an AAV8 capsid having packaged therein a heterologous gene operably linked to regulatory sequences which direct its expression , wherein the heterologous gene encodes factor IX , wherein the AAV8 capsid comprises a vp3 capsid protein having the sequence of aa 203 to 737 of SEQ ID NO: 2 , or a sequence which is at least 95% identical to said sequence of aa 203 to 737 of SEQ ID NO: 2.2. The vector according to claim 1 , further comprising one or more AAV inverted terminal repeat (ITR) sequence from an AAV heterologous to AAV8.3. The vector according to claim 2 , wherein the one or more AAV ITR is from AAV2.4. A composition comprising the vector according to and a pharmaceutically acceptable carrier.5. A host cell containing the vector according to in culture.6. A method for treating hemophilia B claim 1 , said method comprising the step of contacting a cell with a vector according to claim 1 , wherein said vector directs expression of factor IX.7. The method according to claim 6 , wherein said vector is delivered via intravenous administration.8. An adeno-associated virus (AAV)8 viral vector comprising an AAV8 capsid having packaged therein a heterologous gene operably linked to regulatory sequences which direct its expression claim 6 , wherein the heterologous gene encodes ornithine transcarbamylase (OTC) claim 6 , wherein the AAV8 capsid comprises a vp3 capsid protein having the sequence of aa 203 to 737 of SEQ ID NO: 2 claim 6 , or a sequence which is at least 95% identical to said sequence of aa 203 to 737 of SEQ ID NO: 2.9. The vector according to claim 8 , wherein said OTC is human OTC.10. The vector according to claim 8 , further comprising one or more AAV inverted terminal repeat (ITR) ...

ALTERED HOST CELL PATHWAY FOR IMPROVED ETHANOL PRODUCTION

A recombinant yeast cell, fermentation compositions, and methods of use thereof are provided. The recombinant yeast cell includes at least one heterologous nucleic acid encoding one or more polypeptide having phosphoketolase activity; phosphotransacetylase activity; and/or acetylating acetaldehyde dehydrogenase activity, wherein the cell does not include a heterologous modified xylose reductase gene, and wherein the cell is capable of increased biochemical end product production in a fermentation process when compared to a parent yeast cell. 1. A recombinant yeast cell comprising at least one heterologous nucleic acid encoding one or more polypeptide having:i) phosphoketolase activity;ii) phosphotransacetylase activity; and wherein said cell does not comprise a heterologous modified xylose reductase gene,', 'wherein said cell is capable of increased ethanol production from glucose in a fermentation process when compared to the yeast cell without the at least one heterologous nucleic acid, and', {'i': 'Lactobacillus plantarum.', 'wherein the polypeptide having phosphoketolase activity has the amino acid of SEQ ID NO: 57, the polypeptide having acetylating acetaldehyde dehydrogenase activity has the amino acid of SEQ ID NO: 32, and the polypeptide having phophotransacetylase activity is the phophotransacetylase from'}], 'iii) acetylating acetaldehyde dehydrogenase activity,'}2. The recombinant yeast cell of claim 1 , wherein said cell has a reduced NAD-dependant glycerol phosphate dehydrogenase (GPD) activity when compared to a parent yeast cell.3. The recombinant yeast cell of claim 1 , wherein said cell comprises an altered pentose phosphate pathway resulting from one or more heterologously expressed nucleic acid affecting the pentose phosphate pathway.4Saccharomyces cerevisiae.. The recombinant yeast cell of wherein the species of the yeast cell is5. The recombinant yeast cell of claim 1 , wherein said fermentation process is selected from the group consisting of ...

SECRETAGOGUES DERIVED FROM OXALOBACTER FORMIGENES

The present invention relates to a secretagogue compound derived from oxalate degrading bacteria, for use in the treatment of an oxalate related disease and/or oxalate related imbalance in a subject, wherein the administration of the secretagogue results in a reduction of urinary oxalate and/or plasma oxalate in the subject. The invention further relates to a pharmaceutical composition comprising such a secretagogue compound, a method for treating a subject suffering from an oxalate related disease, and to a method for preparing a secretagogue. 1Oxalobacter formigenes. A secretagogue derived from oxalate degrading bacteria , e.g. from , for use in the treatment of an oxalate related disease and/or oxalate related imbalance in a subject , wherein the administration of the secretagogue results in a reduction of urinary oxalate and/or plasma oxalate in the subject.2. The secretagogue for use according to claim 1 , wherein the oxalate related disease is selected from the group consisting of primary hyperoxaluria claim 1 , hyperoxaluria claim 1 , absorptive hyperoxaluria claim 1 , enteric hyperoxaluria claim 1 , idiopathic calcium oxalate kidney stone disease (urolithiasis) claim 1 , vulvodynia claim 1 , oxalosis associated with end-stage renal disease claim 1 , cardiac conductance disorders claim 1 , inflammatory bowel disease claim 1 , Crohn's disease claim 1 , ulcerative colitis claim 1 , and disorders/conditions caused by/associated with gastrointestinal surgery claim 1 , bariatric surgery (surgery for obesity) claim 1 , and/or antibiotic treatment.3. The secretagogue for use according to or claim 1 , comprising an amino acid sequence claim 1 , which has a secretagogue activity and at least 85% identity with an amino acid sequence according to any one of SEQ ID NO: 1-19.4. The secretagogue for use according to claim 3 , comprising an amino acid sequence which has a secretagogue activity and at least 85% identity claim 3 , with an amino acid sequence according to any ...

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Non-viral delivery systems comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A composition comprising a non-viral carrier and a nucleic acid molecule comprising an engineered sequence encoding human ornithine transcarbamylase (hOTCase) and expression control sequences which direct expression of hOTCase in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over at least the mature hOTC sequence of SEQ ID NO: 1 , wherein said hOTC nucleic acid sequence is a DNA sequence of SEQ ID NO: 3.2. The composition according to claim 1 , wherein the expression control sequences further comprise a liver-specific promoter.3. The composition according to claim 2 , wherein the liver-specific promoter is selected from a thyroxin-binding globulin (TBG) promoter or a lymphocyte-specific protein 1 (LSP1) promoter.4. The composition according to claim 1 , wherein the expression cassette further comprises one or more of an intron claim 1 , a Kozak sequence claim 1 , a poly A claim 1 , and a posttranscriptional regulatory elements.5. The composition according to claim 1 , wherein nonviral carrier is a plasmid.6. The composition according to claim 1 , wherein the nucleic acid molecule is formulated in a moiety selected from the group consisting of micelles claim 1 , liposomes claim 1 , cationic lipid-nucleic acid compositions claim 1 , poly-glycan compositions claim 1 , block copolymers and other polymers claim 1 , lipid and/or cholesterol-based-nucleic acid conjugates.7. The composition according to claim 6 , wherein the moiety is a nanoparticle.8. A composition comprising an mRNA according to any one of SEQ ID NO: 10 claim 6 , ...

COMPOSITIONS AND METHODS FOR THE BIOSYNTHESIS OF VANILLIN OR VANILLIN BETA-D-GLUCOSIDE

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express a mutant AROM polypeptide and/or mutant catechol-O-methyltransferase polypeptide alone or in combination with one or more vanillin biosynthetic enzymes or UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce vanillin or vanillin beta-D-glucoside. 1. A method for producing vanillin and/or vanillin beta-D-glucoside comprising(a) providing a recombinant host capable of producing vanillin, wherein said recombinant host harbors a heterologous nucleic acid encoding a mutant Catechol-O-Methyl Transferase (COMT) polypeptide;(b) cultivating said recombinant host for a time sufficient for said recombinant host to produce vanillin and/or vanillin glucoside; and(c) isolating vanillin and/or vanillin glucoside from said recombinant host or from the cultivation supernatant, thereby producing vanillin and/or vanillin beta-D-glucoside.2. The method of claim 1 , wherein the mutant COMT polypeptide is capable of catalyzing methylation of an —OH group of protocatechuic acid claim 1 , wherein said methylation results in generation of at least 4 times more vanillic acid compared to iso-vanillic acid.3. The method of claim 1 , wherein the mutant COMT polypeptide has an amino acid sequence sharing at least 80 percent sequence identity with SEQ ID NO:27 determined over the entire length of SEQ ID NO:27; and which has an amino sequence which differs from SEQ ID NO:27 by at least one amino acid residue.4. The method of claim 1 , wherein the mutant COMT polypeptide comprises 1 to 10 amino acid substitutions in the amino acid sequence set forth in SEQ ID NO:27.5. The method of claim 1 , wherein the mutant COMT polypeptide comprises an amino acid having a lower hydropathy index than leucine at position 198 of SEQ ID NO:27.6. The method of claim 1 , wherein the mutant COMT polypeptide comprises Ala claim 1 , Arg claim 1 , Asn claim 1 , Asp claim 1 , Cys claim ...

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

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 method for producing putrescine , comprising:(i) culturing a recombinant microorganism having putrescine productivity, and(ii) recovering putrescine from the cultured recombinant microorganism or the cell culture,wherein the recombinant microorganism is modified to have enhanced activity of a protein having the amino acid sequence of SEQ ID NO: 21 or 23 as compared to the activity in the same microorganism without the modification.2. The method of claim 1 , wherein the recombinant microorganism further modified to have enhanced ornithine decarboxylase (ODC) activity as compared to the activity in the same microorganism without the modification.3. The method of claim 1 , wherein the recombinant microorganism further modified to have weakened activities of ornithine carbamoyltransferase (ArgF) and a protein involved in glutamate export claim 1 , as compared to the activity in the same microorganism without the modification.4. The method of claim 2 , wherein the ornithine decarboxylase (ODC) comprises the amino acid sequence of SEQ ID NO: 33.5. The method of claim 3 , wherein the ornithine carbamoyltransferase (ArgF) comprises the amino acid sequence of SEQ ID NO: 29 claim 3 , and the protein involved in glutamate export comprises the amino acid sequence of SEQ ID NO: 30.6. The method of claim 1 , wherein the recombinant 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 , as compared to the activities in the same microorganism without the modification.7. ...

ARGININE SUPPLEMENTATION TO IMPROVE EFFICIENCY IN GAS FERMENTING ACETOGENS

The invention provides methods for improving efficiency of fermentation by arginine supplementation, and genetically modified bacterium for use therefor. More particularly the invention provides methods for (i) increasing the production ATP intensive products with arginine supplementation, (ii) increasing utilization of arginine by a C1-fixing bacterium; and (iii) providing C1-fixing bacterium with optimized arginine de-aminase pathways. 1. A method for increasing the production of at least one ATP-intensive product , the method comprising;a. flowing a gaseous C1-containing substrate to a bioreactor containing a culture of a C1-fixing microorganism in a liquid nutrient media; andb. fermenting the culture to produce at least one product;wherein arginine is provided to the culture in excess of the cellular requirement of the culture; and wherein the C1-fixing microorganism comprises an arginine metabolism pathway.2. The method of wherein the arginine metabolism pathway comprises at least one of an arginine deaminase pathway and an arginine decarboxylase pathway claim 1 , wherein the arginine deaminase pathway comprises one or more enzymes selected from the group consisting of arginine deiminase (EC 3.5.3.6) claim 1 , ornithine carbomyltransferase (putrescine carbomyltransferase) (EC 2.1.3.3) and a carbamate kinase (EC 2.7.2.2) and the arginine decarboxylase pathway comprises one or more enzymes selected from the group consisting arginine decarboxylase (EC 4.1.1.19) claim 1 , putative arginine deaminase (EC 3.5.3.12) claim 1 , putrescine carbamoyl transferase (EC 2.1.3.6) and carbamate kinase (EC 2.7.2.2).3. The method of claim 1 , wherein arginine is provided to the culture in an amount ranging from the cellular requirement of the culture to about 1000 times the cellular requirement of the culture.4. The method of claim 1 , wherein arginine is provided to the culture in an amount ranging from 2 times the cellular requirement of the culture to 1000 times the cellular ...

Method for Producing Objective Substance

A method for producing an objective substance such as vanillin and vanillic acid is provided. An objective substance is produced from a carbon source or a precursor of the objective substance by using a microorganism that is able to produce the objective substance, which microorganism has been modified so that the activity of an enzyme involved in SAM cycle (SAM cycle enzyme) is increased. 1. A method for producing an objective substance , the method comprising:producing the objective substance by using a microorganism having an ability to produce the objective substance,wherein the microorganism has been modified so that the activity of an S-adenosylmethionine cycle enzyme is increased as compared with a non-modified microorganism,wherein the S-adenosylmethionine cycle enzyme is selected from the group consisting of methionine adenosyltransferase, methionine synthase, 5,10-methylenetetrahydrofolate reductase, and combinations thereof, andwherein the objective substance is selected from the group consisting of:(X) metabolites the biosynthesis of which requires S-adenosylmethionine;(Y) L-methionine; and(Z) combinations thereof,provided that the objective substance is selected from the metabolites (X) when only the activity of methionine adenosyltransferase is increased.2. The method according to claim 1 , wherein said producing comprises:cultivating the microorganism in a culture medium comprising a carbon source so that the objective substance is produced and accumulates in the culture medium.3. The method according to claim 1 , wherein said producing comprises:converting a precursor of the objective substance into the objective substance by using the microorganism.4. The method according to claim 3 , wherein said using comprises:cultivating the microorganism in a culture medium comprising the precursor so that the objective substance is produced and accumulates in the culture medium.5. The method according to claim 3 , wherein said converting comprises:allowing ...

ACETAMINOPHEN ADDUCTS AND METHODS OF USE THEREOF

The present disclosure relates to acetaminophen protein adducts and methods of diagnosing acetaminophen toxicity using the acetaminophen protein adducts. The present disclosure provides acetaminophen (APAP)-protein adducts and methods of detecting acetaminophen-induced toxicity in a subject using APAP-protein adducts. One aspect of the present disclosure provides an APAP-protein adduct for diagnosing acetaminophen-induced toxicity. According to the present disclosure, the inventors have identified proteins that are modified by N-acetyl-pbenzoquinoneimine (NAPQI) in subjects with acetaminophen-induced toxicity. Non-limiting examples of proteins modified by NAPQI include betaine-homocysteine 5-methyltransferase 1, cytoplasmic aspartate aminotransferase, 1,4-alpha-glucan branching enzyme, formimidoyltransferase-cyclodeaminase, and dystrophin. 1. (canceled)2. (canceled)3. A method for measuring the amount of acetaminophen-protein adduct in a biological sample , the method comprising:a) obtaining a biological sample from a subject; andb) measuring the amount of acetaminophen-protein adduct in the sample by detecting one or more APAP-protein adducts, wherein each APAP-protein adduct comprises a protein modified with NAPQI, and wherein the protein is selected from the group consisting of betaine-homocysteine S-methyltransferase 1, cytoplasmic aspartate aminotransferase, 1,4-alpha-glucan-branching enzyme, formimidoyltransferase-cyclodeaminase, or dystrophin.4. The method of claim 3 , wherein the biological sample is a biological fluid selected from the group consisting of blood claim 3 , plasma claim 3 , serum claim 3 , urine claim 3 , saliva and hair.5. A method of detecting acetaminophen-induced toxicity in a subject claim 3 , the method comprising:a) obtaining a biological sample from the subject;b) measuring the amount of acetaminophen-protein adduct in the sample by detecting one or more APAP-protein adducts, wherein each APAP-protein adduct comprises a protein ...

Messenger rna therapy for the treatment of ornithine transcarbamylase deficiency

The present invention provides, among other things, methods of treating ornithine transcarbamylase (OTC) deficiency, including administering to a subject in need of treatment a composition comprising an mRNA encoding an ornithine transcarbamylase (OTC) protein at an effective dose and an administration interval such that at least one symptom or feature of the OTC deficiency is reduced in intensity, severity, or frequency or has delayed onset. In some embodiments, the mRNA is encapsulated in a liposome comprising one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids.

Multimeric coding nucleic acid and uses thereof

The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends.

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

The present invention relates to a recombinant microorganism for producing putrescine or ornithine, and a method for producing putrescine or ornithine using the same. Specifically, the present invention relates to a microorganism of the genus Corynebacterium capable of producing putrescine or ornithine, in which an activity of the transcriptional regulator of sugar metabolism (SugR) is weakened, an activity of the citrate synthase (GltA) is enhanced, or both are applied; and a method for producing putrescine or ornithine using the same.

COMPOSITIONS AND METHODS FOR TREATING ORNITHINE TRANSCARBAMYLASE DEFICIENCY

The present disclosure provides a modified human OTC protein having improved properties for the treatment of OTC deficiency in a patient. Preferably, the protein of the disclosure is produced from a codon optimized mRNA suitable for administration to a patient suffering from OTC deficiency wherein upon administration of the mRNA to the patient, the protein of the disclosure is expressed in the patient in therapeutically effective amounts to treat OTC deficiency. The present disclosure also provides codon optimized mRNA sequences encoding wild type human OTC comprising a 5′ UTR derived from a gene expressed by for use in treating OTC deficiency in a patient. 1. An ornithine transcarbamylase (OTC) protein comprising the amino acid sequence of SEQ ID NO:4 and having OTC enzymatic activity.2. A polynucleotide encoding the protein of .3. The polynucleotide of comprising an optimized coding region encoding SEQ ID NO: 4.4. The polynucleotide of claim 3 , wherein the polynucleotide is DNA.5. The polynucleotide of claim 3 , wherein the polynucleotide is an mRNA.6. (canceled)7. The polynucleotide of comprising a 3′ poly A tail having from about 60 sequential adenine nucleotides to about 125 sequential adenine nucleotides.8. (canceled)9. The polynucleotide of comprising a non-human 5′ untranslated region (5′UTR).10Arabidopsis thaliana.. The polynucleotide of claim 9 , wherein the 5′UTR is derived from a gene expressed by11. The polynucleotide of claim 10 , wherein the 5′ UTR comprises a sequence selected from SEQ ID NO: 6 claim 10 , SEQ ID NOS: 125-127 and SEQ ID NOS: 227-247.12. (canceled)13. The polynucleotide of comprising a 3′ untranslated region (3′ UTR) selected from the group consisting of: SEQ ID NOS: 16-22.14. (canceled)15. The polynucleotide of comprising a Kozak sequence of SEQ ID NO: 23 or a partial Kozak sequence of SEQ ID NO: 24.16. The polynucleotide of comprising a 5′ cap.18. The polynucleotide of claim 4 , wherein the optimized coding region comprises SEQ ID ...

METHODS OF ALTERING SEED WEIGHT AND SEED OIL CONTENT BY MANIPULATING ALPHA-CARBOXYL-TRANSFERASE (A-CT) ACTIVITY VIA CARBOXYL-TRANSFERASE INTERACTION (CTI) PROTEIN EXPRESSION

A method of altering fatty acid and/or triacylglycerol production in plants and/or algae is provided. The method includes altering activity levels of alpha-carboxyltransferase (α-CT), a catalytic subunit of acetyl-CoA carboxylase (ACCase) by modulating an expression of at least one carboxyltransferase interaction (CTI) gene encoding at least one CTI protein. 1. A method of altering fatty acid and/or triacylglyerol production in plants and/or algae , comprising altering activity levels of alpha-carboxyltransferase (α-CT) , wherein α-CT comprises a catalytic subunit of acetyl-CoA carboxylase (ACCase).2. The method of claim 1 , wherein altering activity levels of α-CT comprises altering intracellular concentrations of one or more carboxyl transferase interactor (CTI) proteins claim 1 , wherein the one or more CTI proteins inhibit activity levels of α-CT.3. The method of claim 2 , wherein altering intracellular concentrations of the one or more CTI proteins further comprises altering expression of one or more carboxyl transferase interactor (CTI) genes.4. The method of claim 3 , wherein the one or more CTI genes comprise genes and gene orthologs of CTI1 claim 3 , CTI2 claim 3 , and CTI3 claim 3 , or artificial genes containing essential CTI motifs.5. The method of claim 3 , wherein the one or more CTI genes comprise from about 70% to about 100% sequence identity to a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1 claim 3 , 3 claim 3 , 5 claim 3 , and 34-54 or a complement thereof claim 3 , or wherein the one or more CTI genes encode a CTI protein with a polypeptide sequence ranging from about 70% to about 100% sequence identity to a polypeptide sequence selected from the group consisting of SEQ ID NOs: 2 claim 3 , 4 claim 3 , 6 claim 3 , 7-28 claim 3 , and 60-92.6. (canceled)7. The method of claim 3 , wherein altering fatty acid and/or triacylglycerol production in plants comprises increasing fatty acid and/or triacylglycerol production in ...

Compositions and methods for the biosynthesis of vanillin or vanillin beta-d-glucoside

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express a mutant AROM polypeptide and/or mutant catechol-O-methyltransferase polypeptide alone or in combination with one or more vanillin biosynthetic enzymes or UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce vanillin or vanillin beta-D-glucoside.

ARGININE SUPPLEMENTATION TO IMPROVE EFFICIENCY IN GAS FERMENTING ACETOGENS

The invention provides methods for improving efficiency of fermentation by arginine supplementation, and genetically modified bacterium for use therefor. More particularly the invention provides methods for (i) increasing the production ATP intensive products with arginine supplementation, (ii) increasing utilization of arginine by a C1-fixing bacterium; and (iii) providing C1-fixing bacterium with optimized arginine de-aminase pathways. 1. A method for increasing the production of at least one ATP-intensive product , the method comprising;a. flowing a gaseous C1-containing substrate to a bioreactor containing a culture of a C1-fixing microorganism in a liquid nutrient media; andb. fermenting the culture to produce at least one product;wherein arginine is provided to the culture in excess of the cellular requirement of the culture; and wherein the C1-fixing microorganism comprises an arginine metabolism pathway.2. The method of claim 1 , wherein arginine is provided to the culture in an amount ranging from 2 times the cellular requirement of the culture to 1000 times the cellular requirement of the culture.3. The method of claim 1 , wherein the cellular requirement of the culture is about 0.012 g of arginine per gram of cellular biomass.4. The method of claim 1 , wherein the doubling time of the culture is decreased by at least 10% when compared to a culture where arginine is not provided in excess of the cellular requirement of the culture.5. The method of claim 4 , wherein the doubling time of the culture is at decreased by at least 50% when compared to a culture where arginine is not provided in excess of the cellular requirement of the culture.6. The method of claim 1 , wherein selectivity to the ATP-intensive product is increased when compared to a culture where arginine is not provided in excess of the cellular requirement of the culture.7. The method of claim 1 , wherein productivity of the ATP-intensive product is at least 10% greater when compared to a ...

Stable Gene Transfer to Proliferating Cells

Provided herein are methods for facilitating or inducing stable transgene integration and expression in a proliferating cell, comprising administering to the cell (i) a recombinant AAV (rAAV) vector comprising the transgene flanked by transposon-derived inverted terminal repeat sequences, which sequences are in turn flanked by AAV-derived inverted terminal repeat regions, and (ii) a source of a transposase that recognises said transposon-derived inverted terminal repeat sequences and directs the genomic integration of the transgene into the genome of the proliferating cell. Also provided are methods and transgene delivery systems for the treatment or prevention of diseases affecting, associated with or characterised by proliferating cells, including paediatric liver diseases, bone marrow diseases and cancer. 1. A method for stably integrating a transgene into the genome of a proliferating cell and/or inducing stable transgene expression in a proliferating cell , the method comprising administering to the cell: (i) a recombinant AAV (rAAV) vector comprising the transgene flanked by transposon-derived inverted terminal repeat sequences , which sequences are in turn flanked by AAV-derived inverted terminal repeat regions; and (ii) a source of a transposase that recognises said transposon-derived inverted terminal repeat sequences and directs the genomic integration of the transgene into the genome of the proliferating cell.2. A method according to claim 1 , wherein the genomic integration of the transgene into the genome of the proliferating cell facilitates or induces the stable transgene expression.34-. (canceled)5. A method according to claim 1 , wherein the transgene and flanking transposon-derived inverted terminal repeat sequences form a transposon-transgene cassette claim 1 , optionally comprising one or more further sequences or genetic elements including claim 1 , for example claim 1 , a promoter claim 1 , enhancer claim 1 , post-regulatory element and/or ...

A microorganism of genus corynebacterium having an ability to produce l-arginine and a method for producing l-arginine using the same

The present invention relates to a microorganism of the genus Corynebacterium having an ability to produce L-arginine, and a method of producing L-arginine using the same.

RICE SERINE HYDROXYMETHYLTRANSFERASE CODED GENE OsSHM4 MUTANTS AND APPLICATION THEREOF

The present invention discloses rice serine hydroxymethyltransferase coded gene OsSHM4 mutants and application thereof. The rice serine hydroxymethyltransferase coded gene mutants are obtained in a manner that T at a 461st position of a CDS sequence region of a wild type rice OsSHM4 gene is mutated to C, so that coded amino acids thereof are mutated from leucine to proline. A CDS sequence of the wild type rice OsSHM4 gene is shown in SEQ ID NO. 1. After mutation of serine hydroxymethyltransferase coded genes OsSHM4, under a field experiment condition, S and Se content of rice shoots is remarkably improved without influencing biomass of rice. After wild type serine hydroxymethyltransferase coded genes OsSHM4 are genetically modified to complement the mutants, S and Se content of shoots of complemented strains is restored to that of wild type rice water planting. 1. A rice serine hydroxymethyltransferase coded mutant gene , wherein the rice serine hydroxymethyltransferase mutant gene has a C instead of a T at a 461st position of the coding sequence (CDS) region of a wild type rice OsSHM4 gene , so as to encode a proline instead of a leucine at the corresponding coded amino acid thereof.2. The mutant gene of claim 1 , wherein the CDS sequence of the wild type rice OsSHM4 gene is set forth as SEQ ID NO. 1.3. A protein encoded by the rice serine hydroxymethyltransferase coded gene mutant according to .4. The protein of having an amino acid sequence set forth as SEQ ID NO.4.5. An expression vector comprising the rice serine hydroxymethyltransferase coded mutant gene according to .6. A host cell comprising the rice serine hydroxymethyltransferase coded mutant gene according to .7. A method for enhancing an absorbing capacity of rice roots to sulfate and improving total sulfur concentration of rice shoots claim 1 , for enhancing an absorbing capacity of roots to selenate and improving total selenium concentration of rice shoots claim 1 , or for improving total S and Se ...

mRNA FOR USE IN TREATMENT OF HUMAN GENETIC DISEASES

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification conferring stability to the RNA, as well as related methods, are disclosed. 1. A composition for delivery of mRNA for the expression of a protein in a target cell , comprising at least one mRNA molecule comprising a sequence encoding a protein of interest and a 3′ untranslated region (3′-UTR) comprising a sequence encoding a portion of human growth hormone (hGH).2. (canceled)3. The composition of claim 1 , wherein the mRNA molecule comprises more than one modification which confers stability to the mRNA molecule.4. The composition of claim 1 , wherein the mRNA molecule comprises a modification of the 5′ untranslated region.5. The composition of claim 4 , wherein said modification comprises a partial sequence of a CMV immediate-early 1 (IE 1) gene.6. The composition of claim 5 , wherein said partial sequence of the CMV immediate-early 1 (IEI) gene comprises SEQ ID NO: 2 or SEQ ID NO: 1.7. The composition of claim 1 , wherein the mRNA comprises the inclusion of a poly A tail.8. The composition of claim 1 , wherein the mRNA comprises the inclusion of a Cap 1 structure.910-. (canceled)11. The composition of claim 1 , wherein the sequence encoding a portion of human growth hormone (hGH) comprises SEQ ID NO: 3.12. (canceled)13. The composition of claim 1 , wherein the mRNA encodes ornithine carbamoyl transferase.14. The composition of claim 1 , wherein the mRNA encodes alpha galactosidase.15. The composition of claim 1 , wherein the mRNA encodes erythropoietin.16. A method of treating a subject deficient in a protein claim 1 , comprising administering a composition comprising an mRNA and a transfer vehicle claim 1 , wherein the mRNA encodes a protein corresponding to the protein which is deficient in the subject claim 1 , and wherein the mRNA comprises comprises a 3′ untranslated region (3′-UTR) comprising a sequence ...

MITOCHONDRIAL PROTEINS CONSTRUCTS AND USES THEREOF

Disclosed are novel fusion protein constructs comprising a functional mitochondrial protein, that can enter mitochondria within intact cells. Further disclosed are methods of treating mitochondrial disorders by the disclosed fusion proteins and compositions therefor. 1. A fusion protein comprising an HIV-1 transactivator of transcription (TAT) domain fused to a human pyruvate dehydrogenase E1 component alpha subunit (PDHE1a) and a human mitochondria targeting sequence (MTS) , wherein the MTS is situated between the TAT domain and the PDHE1a , the PDHE1a is C-terminal to the MTS , and the MTS is heterologous to the PDHE1a.2. The fusion protein of claim 1 , wherein the MTS is a human citrate synthase (CS) MTS.3. The fusion protein of claim 2 , wherein the MTS comprises the amino acid sequence of SEQ ID NO. 23.4. The fusion protein of claim 1 , further comprising a linker covalently linking the TAT domain to the MTS.5. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the fusion protein according to as an active ingredient.6. A method for introducing a human pyruvate dehydrogenase E1 component alpha subunit (PDHE1a) into a subject claim 1 , comprising administering to the subject the fusion protein of .7. A method for treating or alleviating a mitochondrial disorder claim 1 , comprising administering to a human subject in need thereof the fusion protein of .8. The method of claim 7 , wherein the fusion protein is intravenously administered to the subject. The present application is a continuation application of U.S. Ser. No. 14/784,253, filed Oct. 13, 2015, which is US national stage application of International Application PCT/IL2014/050354, filed Apr. 10, 2014, which is a continuation-in-part of U.S. Ser. No. 14/034,224, filed Sep. 23, 2013, now U.S. Pat. No. 8,912,147, issued Dec. 16, 2014, which claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Ser. No. 61/869,981, filed Aug. 26, 2013, and, which claims the ...

MULTIMERIC CODING NUCLEIC ACID AND USES THEREOF

The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends. 142-. (canceled)43. A method of delivering a multimeric coding nucleic acid (MCNA) for in vivo protein production , comprising administering to a subject in need of delivery a MCNA , wherein the MCNA comprises two messenger RNAs (mRNAs) linked at 3′ ends , via stable linkage , such that the multimeric coding nucleic acid has two 5′ ends , and wherein the stable linkage is an oligonucleotide bridge comprising an internal 3′-to-3′ inverted phosphodiester linkage.4447-. (canceled)48. The method of claim 43 , wherein each of the mRNAs encodes a protein of interest.49. The method of claim 48 , wherein each of the mRNAs encodes a same protein.50. The method of claim 48 , wherein each of the mRNAs encodes a distinct protein.51. The method of claim 43 , wherein the mRNAs comprise a 3′ UTR.52. The method of claim 51 , wherein the 3′ UTR comprises a plurality of multi-A segments with spacers in between.53. The method of claim 43 , wherein the oligonucleotide bridge comprises nucleosides selected from the group consisting of 2′-OMe-A claim 43 , 2′-OMe-G claim 43 , 2′-OMe-C claim 43 , 2′-OMe-U claim 43 , 2′-F-A claim 43 , 2′-F-G claim 43 , 2′-F-C claim 43 , 2′-F-U claim 43 , LNA-A claim 43 , LNA-G claim 43 , LNA-C claim 43 , LNA-U claim 43 , N6-methyl-adenosine claim 43 , 2-thiouridine (2sU) claim 43 , 5-methyl-cytidine (5mC) claim 43 , pseudouridine (ψU) claim 43 , and 1-methyl-pseudouridine.54. The method of claim 43 , wherein the mRNAs comprise one or more modified nucleosides.55. The method of claim 54 , wherein the modified nucleosides are selected from the group consisting of 2′-OMe-A claim 54 , 2′-OMe-G claim 54 , 2′- ...

PRODUCTION OF MALONYL-COA DERIVED PRODUCTS VIA ANAEROBIC PATHWAYS

The present invention provides for novel metabolic pathways to convert biomass and other carbohydrate sources to malonyl-CoA derived products, such as hydrocarbons and other bioproducts, under anaerobic conditions and with the net production of ATP. More specifically, the invention provides for a recombinant microorganism comprising one or more native and/or heterologous enzymes that function in one or more engineered metabolic pathways to achieve conversion of a carbohydrate source to, e.g., long-chain hydrocarbons and hydrocarbon derivatives, wherein the one or more native and/or heterologous enzymes is activated, upregulated, downregulated, or deleted. The invention also provides for processes to convert biomass to malonyl-CoA derived products which comprise contacting a carbohydrate source with a recombinant microorganism of the invention. 1132-. (canceled)133. A recombinant yeast microorganism comprisingone or more engineered metabolic pathways to convert a carbohydrate source to a hydrocarbon, (a) the conversion of phosphoenolpyruvate to oxaloacetate by a phosphoenolpyruvate carboxykinase and', '(b) the conversion of oxaloacetate and acetyl-CoA to malonyl-CoA and pyruvate by a heterologous transcarboxylase Enzyme Commission Number 2.1.3.1;, 'wherein the one or more engineered metabolic pathways comprises'}wherein the one or more engineered metabolic pathways further comprises downregulation or deletion of native pyruvate decarboxylase, and wherein the one or more engineered metabolic pathway further comprises a heterologous pyruvate formate lyase, pyruvate dehydrogenase, pyruvate:ferredoxin oxidoreductase or pyruvate:NADP+oxidoreductase.134. The recombinant microorganism of claim 133 , wherein the conversion of a carbohydrate source to a hydrocarbon is under anaerobic or microaerophilic conditions.135. The recombinant microorganism of claim 133 , wherein said hydrocarbon is selected from the group consisting of:(a) an alkane;(b) an alkene;(c) an alkyne;(d) a ...

ENZYMATIC SYSTEMS FOR CARBON FIXATION AND METHODS OF GENERATING SAME

A system for carbon fixation is provided. The system comprises enzymes which catalyze reactions of a carbon fixation pathway, wherein at least one of the reactions of the carbon fixation pathway is a carboxylation reaction, wherein products of the reactions of the carbon fixation pathway comprise oxaloacetate and malonyl-CoA, wherein an enzyme which performs the carboxylation reaction is selected from the group consisting of phophoenolpyruvate (PEP) carboxlase, pyruvate carboxylase and acetyl-CoA carboxylase and wherein an export product of the carbon fixation pathway is glyoxylate. Additional carbon fixation pathways are also provided and methods of generating same. 1. A system for carbon fixation , comprising an electron donor and enzymes which catalyze reactions of a carbon fixation pathway , wherein all the carboxylation reactions of the carbon fixation pathway utilize:(i) phophoenolpyruvate (PEP) carboxylase and acetyl-CoA carboxylase; or(ii) phophoenolpyruvate (PEP) carboxylase; or(iii) pyruvate carboxylase and acetyl-CoA carboxylase; or(iv) pyruvate carboxylasewherein products of said reactions of the carbon fixation pathway comprise oxaloacetate and malonyl-CoA, and wherein an additional product of the carbon fixation pathway is glyoxylate, wherein an in-organic carbon is introduced into a substrate to become a carboxylic acid group during said carboylation reactions.2. The system of claim 1 , wherein said glyoxylate is an export product.3. The system of claim 1 , wherein an export product of the carbon fixation pathway is pyruvate.4. The system of claim 1 , wherein the enzymes of the carbon fixation pathway generate more than 0.3 μmol glyceraldehyde-3-phosphate/min/mg.5. The system of claim 1 , wherein said enzyme which performs said carboxylation enzyme is PEP carboxylase.6. The system of claim 1 , wherein at least two of said reactions of the carbon fixation pathway are carboxylation reactions.7. The system of claim 1 , wherein one of said reactions of ...

ADENO-ASSOCIATED VIRUS (AAV) SEROTYPE 8 SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. 1(a) a molecule encoding the vp1 capsid protein having a sequence of amino acids 1 to 738 of SEQ ID NO: 2, or a sequence which is at least 95% identical to the full length of amino acids 1 to 738 of SEQ ID NO: 2;(b) a functional rep gene;(c) a nucleic acid molecule comprising at least one AAV inverted terminal repeat (ITR) and a non-AAV nucleic acid sequence encoding a gene product operably linked to sequences which direct expression of the product in a host cell; and(d) sufficient helper functions to permit packaging of the minigene into the AAV capsid protein under conditions which permit packaging of the minigene into the AAV capsid.. A method of generating a recombinant adeno-associated virus (AAV) comprising culturing a host cell containing: This application is a continuation of U.S. patent application Ser. No. 16/142,921, filed Sep. 26, 2018, which is a continuation of U.S. patent application Ser. No. 15/298,760, filed Oct. 20, 2016, now U.S. Pat. No. 10,266,846, issued Apr. 23, 2019, which is a divisional of U.S. patent application Ser. No. 14/598,462, filed Jan. 16, 2015, now U.S. Pat. No. 9,493,788, issued Nov. 15, 2016, which is a divisional of U.S. patent application Ser. No. 11/981,022, filed Oct. 31, 2007, now U.S. Pat. No. 8,962,330, issued Feb. 24, 2015, which is a continuation of U.S. patent application Ser. No. 11/899,500, filed Sep. 6, 2007, now U.S. Pat. No. 7,790,449, issued Sep. 7, 2010, which is a continuation of U.S. patent application Ser. No. 10/423,704, filed Apr. 25, 2003, now U.S. Pat. No. 7,282,199, issued Oct. 16, 2007, which is a continuation-in-part of International Patent Application No. PCT/US02/33630, filed Nov. 12, 2002, which claims the benefit under 35 USC 119(e) of U.S. Provisional Patent Application No. 60/386,122, ...

ADENO-ASSOCIATED VIRUS (AAV) SEROTYPE 8 SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. 1. (canceled)2. A host cell comprising a recombinant nucleic acid molecule:(a) encoding an AAV8 vp1 capsid protein having a sequence comprising amino acids 1 to 738 of SEQ ID NO: 2; or(b) comprising nucleotides 2121 to 4334 of SEQ ID NO: 1, or a nucleotide sequence at least 99% identical to nucleotides 2121 to 4334 of SEQ ID NO: 1, a functional rep gene, a minigene comprising AAV inverted terminal repeats (ITRs) and a transgene, and sufficient helper functions to permit packaging of the minigene into the AAV capsid, wherein the transgene encodes factor IX.3. The host cell according to claim 1 , wherein said recombinant nucleic acid molecule comprises a sequence selected from:vp1, nt 2121 to 4334;vp2, nt 2532 to 4334; orvp3, nt 2730 to 4334 of SEQ ID NO: 1.4. The host cell according to claim 1 , wherein said recombinant nucleic acid molecule is a plasmid.5. The host cell according to claim 1 , which further comprises a nucleic acid sequence(a) encoding an AAV8 vp2 capsid protein having a sequence comprising amino acids 138 to 738 of SEQ ID NO: 2; or(b) comprising nucleotides 2532 to 4334 of SEQ ID NO: 1, or a nucleotide sequence at least 99% identical to nucleotides 2532 to 4334 of SEQ ID NO: 1.6. The host cell according to claim 1 , which further comprises a nucleic acid sequence(a) encoding an AAV8 vp3 capsid protein having a sequence comprising amino acids 204 to 738 of SEQ ID NO: 2; or(b) comprising nucleotides 2730 to 4334 of SEQ ID NO: 1, or a nucleotide sequence at least 99% identical to nucleotides 2730 to 4334 of SEQ ID NO: 1.7. An AAV8 vector comprising an AAV8 capsid claim 1 , said capsid having packaged therein a nucleic acid molecule comprising AAV inverted terminal repeat sequences and a gene encoding factor IX claim 1 , which gene is operably ...

ADENO-ASSOCIATED VIRUS (AAV) SEROTYPE 8 SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. 1. An adeno-associated virus (AAV)8 viral vector comprising an AAV8 capsid having packaged therein a heterologous gene operably linked to regulatory sequences which direct its expression, wherein the heterologous gene encodes factor IX. This application is a continuation of U.S. patent application Ser. No. 15/084,615, filed Mar. 30, 2016, which is a divisional of U.S. patent application Ser. No. 14/598,462, filed Jan. 16, 2015, now U.S. Pat. No. 9,493,788, issued Nov. 15, 2016, which is a divisional of U.S. patent application Ser. No. 11/981,022, filed Oct. 31, 2007, now U.S. Pat. No. 8,962,330, issued Feb. 24, 2015, which is a continuation of U.S. patent application Ser. No. 11/899,500, filed Sep. 6, 2007, now U.S. Pat. No. 7,790,449, issued Sep. 7, 2010, which is a continuation of U.S. patent application Ser. No. 10/423,704, filed Apr. 25, 2003, now U.S. Pat. No. 7,282,199, issued Oct. 16, 2007, which is a continuation-in-part of International Patent Application No. PCT/US02/33630, filed Nov. 12, 2002, which claims the benefit under 35 USC 119(e) of U.S. Provisional Patent Application No. 60/386,122, filed Jun. 5, 2002, U.S. Provisional Patent Application No. 60/377,133, filed May 1, 2002, and U.S. Provisional Patent Application No. 60/341,151, filed Dec. 17, 2001, which applications are incorporated herein by reference in their entireties.This invention was made with government support under Grant No. P30 DK 47757-09 awarded by the National Institute of Diabetes and Digestive and Kidney Diseases, and Grant No. P01 HL 59407-03 awarded by the National Heart, Lung, and Blood Institute. The government has certain rights in the invention.The Sequence Listing material filed in electronic form herewith is hereby incorporated by reference. This file is labeled ...

POLYNUCLEOTIDES ENCODING ORNITHINE TRANSCARBAMYLASE FOR THE TREATMENT OF UREA CYCLE DISORDERS

This disclosure relates mRNA therapy for the treatment of ornithine transcarbamylase deficiency (OTCD). mRNAs for use in the invention, when administered in vivo, encode human ornithine transcarbamylase (OTC), isoforms thereof, functional fragments thereof, and fusion proteins comprising OTC. mRNAs of the invention are preferably encapsulated in lipid nanoparticles (LNPs) to effect efficient delivery to cells and/or tissues in subjects, when administered thereto. mRNA therapies of the invention increase and/or restore deficient levels of OTC expression and/or activity in subjects. mRNA therapies of the invention further decrease levels of toxic ammonia associated with deficient OTC activity in subjects. 1. A pharmaceutical composition comprising an mRNA comprising an open reading frame (ORF) encoding an ornithine transcarbamylase (OTC) polypeptide , wherein the composition when administered as a single intravenous dose to a human subject in need thereof is sufficient to:(i) increase the level of OTC activity in liver tissue to within at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of normal OTC activity level for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks post-administration;(ii) increase the level of OTC activity in liver tissue at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 20-fold, or at least 50-fold compared to the subject's baseline OTC activity level or a reference OTC activity level in a human subject having ornithine transcarbamylase deficiency (OTCD) for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at ...

ACETAMINOPHEN ADDUCTS AND METHODS OF USE THEREOF

The present disclosure relates to acetaminophen protein adducts and methods of diagnosing acetaminophen toxicity using the acetaminophen protein adducts. 1. A method for measuring the amount of acetaminophen-protein adduct in a biological sample , the method comprising:a) obtaining a biological sample from a subject having or suspected of having acetaminophen toxicity; andb) measuring the total amount of acetaminophen-protein adduct in the sample, wherein the acetaminophen-protein adducts are detected using high performance liquid chromatography (HPLC) or an immunoassay comprising an antibody with specific for NAPQI bound to a protein through a 3-(cystein-S-yl) APAP (3-Cys-A)-protein linkage.2. The method of claim 1 , wherein the biological sample is a biological fluid selected from the group consisting of blood claim 1 , plasma claim 1 , serum claim 1 , urine claim 1 , saliva and hair.3. The method of claim 1 , wherein the total acetaminophen-protein adducts measured include betaine-homocysteine S-methyltransferase 1 claim 1 , cytoplasmic aspartate aminotransferase claim 1 , 1 claim 1 ,4-alpha-glucan-branching enzyme claim 1 , formimidoyltransferase-cyclodeaminase claim 1 , or dystrophin.4. The method of claim 1 , wherein the total acetaminophen-protein adducts are detected using HPLC with electrochemical detection (HPLC-ECD).5. A method of detecting acetaminophen-induced toxicity in a subject having or suspected of having acetaminophen-induced toxicity claim 1 , the method comprising:a) obtaining a biological sample from the subject;b) measuring the total amount of acetaminophen-protein adduct in the sample, wherein the acetaminophen-protein adducts are detected using high performance liquid chromatography (HPLC) or an immunoassay comprising an antibody with specific for NAPQI bound to a protein through a 3-(cystein-S-yl) APAP (3-Cys-A)-protein linkage; andc) comparing the amount of acetaminophen-protein adduct in the sample to a reference value, wherein a greater ...

MULTIMERIC CODING NUCLEIC ACID AND USES THEREOF

The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends. 1. A multimeric coding nucleic acid (MCNA) comprising two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends.23-. (canceled)4. The MCNA of claim 1 , wherein each of the two or more encoding polynucleotides is a synthetic polydeoxyribonucleotide or a polyribonucleotide.5. The MCNA of claim 1 , wherein each of the two or more encoding polynucleotides encodes a protein of interest.6. The MCNA of claim 5 , wherein each of the two or more encoding polynucleotides encodes a same protein.7. The MCNA of claim 5 , wherein each of the two or more encoding polynucleotides encodes a distinct protein.8. The MCNA of claim 1 , wherein the compound comprises three or more encoding polynucleotides.911-. (canceled)1211. The MCNA of claim claim 1 , wherein the one or more of the encoding polynucleotides comprise a 3′ UTR.13. (canceled)14. The MCNA of claim 12 , wherein the 3′ UTR comprises a plurality of multi-A segments with spacers in between.1525-. (canceled)26. The MCNA of claim 1 , wherein the 3′ ends of the two or more encoding polynucleotides are linked via an oligonucleotide bridge comprising a 3′-3′ inverted phosphodiester linkage.27. The MCNA of claim 26 , wherein the nucleotides comprising the oligonucleotide bridge are selected from the group consisting of 2′-OMe-A claim 26 , 2′-OMe-G claim 26 , 2′-OMe-C claim 26 , 2′-OMe-U claim 26 , 2′-F-A claim 26 , 2′-F-G claim 26 , 2′-F-C claim 26 , 2′-F-U claim 26 , LNA-A claim 26 , LNA-G claim 26 , LNA-C claim 26 , LNA-U claim 26 , N6-methyl-adenosine claim 26 , 2-thiouridine (2sU) claim 26 , 5- ...

ORNITHINE TRANSCARBAMYLASE CODING POLYRIBONUCLEOTIDES AND FORMULATIONS THEREOF

In certain aspects, the disclosure relates to compositions comprising modified Ornithine transcarbamylase (OTC) polyribonucleotides and methods of use. 1. A modified polyribonucleotide comprising a primary sequence which that is at least 95% identical to SEQ ID NO: 4 and which encodes an ornithine transcarbamylase (OTC) protein , wherein the modified polyribonucleotide contains a combination of unmodified and modified ribonucleotides , wherein 30-50% of the uridines are analogs of uridine and 5-30% of the cytidines are analogs of cytidine.2. (canceled)3. (canceled)4. The modified polyribonucleotide of claim 1 , wherein the modified polyribonucleotide contains a combination of unmodified and modified ribonucleotides claim 1 , wherein 30-45% of the uridines are analogs of uridine and 5-20% of the cytidines are analogs of cytidine; orwherein the modified polyribonucleotide is made using an input mixture of ribonucleotides, wherein 30-45% of the uridines are analogs of uridine and 5-20% of the cytidines are analogs of cytidine.5. (canceled)6. A modified polyribonucleotide comprising a primary sequence at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 21-27 claim 1 , wherein the modified polyribonucleotide contains a combination of unmodified and modified ribonucleotides claim 1 , wherein 5-50% of the uridines are analogs of uridine and 5-50% of the cytidines are analogs of cytidine.7. The modified polyribonucleotide of claim 6 , wherein 25-50% of uridines are analogs of uridine and 5-30% of cytidines are analogs of cytidine.8. (canceled)9. (canceled)10. (canceled)11. (canceled)12. The modified polyribonucleotide of claim 1 , wherein the modified polyribonucleotide is codon-optimized for expression in mammalian cells.13. (canceled)14. The modified polyribonucleotide of claim 1 , wherein the modified polyribonucleotide further comprises a 3′ UTR claim 1 , a 5′ UTR claim 1 , or a 3′ UTR and a 5′ UTR claim 1 , and wherein the UTR(s) aid( ...

FUSION PROTEINS

Provided are fusion target-binding proteins comprising a target binding moiety, an intracellular signalling region and a domain that promotes synthesis of arginine or an arginine precursor. The domain may be an enzyme domain such as an argininosuccinate synthase (ASS-1) enzyme domain, or an ornithine transcarbamylase (OTC) enzyme domain. Also provided are cells comprising such a fusion target-binding protein (for example cells that express the fusion target-binding protein), and nucleic acids encoding such fusion target-binding proteins. The invention also provides fusion target-binding proteins comprising a target binding moiety, an intracellular signalling region and a domain that promotes synthesis of tryptophan or a tryptophan precursor. Pharmaceutical compositions, medical uses, and methods of treatment, all using the fusion target-binding proteins, cells, or nucleic acids are disclosed. The proteins, cells, nucleic acids and pharmaceutical compositions may be used in the prevention and/or treatment of cancer, such as neuroblastoma or acute myeloid leukaemia. 1. A fusion target-binding protein comprising a target binding moiety , an intracellular signalling region and a domain that promotes synthesis of arginine or an arginine precursor.2. The fusion target-binding protein according to claim 1 , wherein the domain that promotes synthesis of arginine or an arginine precursor comprises an enzyme domain.3. The fusion target-binding protein according to claim 2 , wherein the enzyme domain is selected from the group consisting of: an ASS-1 domain; an OTC domain; an ASL domain; an OCD1 domain; an ArgG domain; an ArgH domain; and an ArgF domain.4. The fusion target-binding protein according to claim 3 , wherein the ASS-1 domain comprises the amino acid sequence of SEQ ID NO. 1.5. The fusion target-binding protein according to claim 3 , wherein the OTC domain comprises the amino acid sequence of SEQ ID NO. 2 claim 3 , the ASL domain comprises the amino acid sequence ...

DNA ENCODING NOVEL ENZYME HAVING D-SERINE SYNTHASE ACTIVITY, METHOD OF PRODUCING THE ENZYME AND METHOD OF PRODUCING D-SERINE BY USING THE SAME

This invention relates to DNA encoding a novel enzyme having activity of synthesizing D-serine from formaldehyde and glycine, recombinant DNA constructed by integrating such DNA into a vector, a transformant transformed with the recombinant DNA, and a method for producing D-serine from formaldehyde and glycine with the use of the enzyme. 115-. (canceled)16. An isolated protein comprising:(a) the amino acid sequence set forth in SEQ ID NO: 6; or(b) the amino acid sequence set forth in SEQ ID NO: 6 except that from one to five amino acid residues of the amino acid sequence of SEQ ID NO: 6 are substituted as follows:a substitution at position 6 to Met or Ile,a substitution at position 10 to Val or Ala,a substitution at position 14 to Pro or Ala,a substitution at position 17 to Pro or Ala,a substitution at position 22 to Ala or Ser,a substitution at position 23 to Arg or Ser,a substitution at position 24 to Val or Ile,a substitution at position 34 to Pro, Ala or Thr,a substitution at position 35 to Pro or Ala,a substitution at position 51 to Glu or Asp,a substitution at position 68 to Leu or Arg,a substitution at position 87 to Leu or Val,a substitution at position 95 to Arg or Thr,a substitution at position 107 to Ala or Gln,a substitution at position 110 to Ala or Arg,a substitution at position 113 to Ala or Gly,a substitution at position 118 to Ala or Thr,a substitution at position 121st to Ile, Met or Leu,a substitution at position 129 to Ala, Gln or Glu,a substitution at position 133 to Gln or Arg,a substitution at position 134 to Ile or Leu,a substitution at position 136 to His, Ala or Gln,a substitution at position 139 to Ala or Thr,a substitution at position 140 to Ala, Gln or Arg,a substitution at position 165 to Thr or Leu,a substitution at position 175 to Asp or Ala,a substitution at position 179 to Leu or Val,a substitution at position 180 to Asn or Thr,a substitution at position 182 to Val or Ala,a substitution at position 194 to Tyr or Leu,a substitution ...

LIVE ATTENUATED CATFISH VACCINE AND METHOD OF MAKING

Live attenuated bacteria vaccines against enteric septicemia of fish, especially catfish, and methods related to the same. Mutant strains of the bacteria (a pathogenic bacterial strain of Enterobacteriaceae) are provided. The mutant bacteria (or other pathogenic bacterial strain of Enterobacteriaceae) contain one or more gene deletions or disruptions that result in less virulent bacterial strains as live attenuated vaccine compositions against virulent wild-type bacteria (or other pathogenic bacterial strain of Enterobacteriaceae). The mutant strains showing the best immunological protection and safety as a vaccine are the triple mutants ESC-NDKL1 (ΔgcvPΔsdhCΔfrdA) strain and ESC-NDKL2 (ΔgcvPΔsdhCΔmdh) strain, with the ESC-NDKL1 strain providing the greatest safety and efficacy of these two triple mutants. 1Edwardsiella ictaluriEdwardsiella ictaluri. A composition for providing immunological protection from an enteric septicemia caused by , said composition comprises a live attenuated strain of comprising gene disrupting mutations in genes coding for three proteins in the group consisting of glycine cleavage system (gcvP) , succinate dehydrogenase (sdhC) , malate dehydrogenase (mdh) , and fumarate reductase (frdA).2. The composition of claim 1 , wherein the gene disrupting mutations in genes coding for three proteins are glycine cleavage system (gcvP) claim 1 , succinate dehydrogenase (sdhC) claim 1 , and fumarate reductase (frdA).3. The composition of claim 1 , wherein the gene disrupting mutations in genes coding for three proteins are glycine cleavage system (gcvP) claim 1 , succinate dehydrogenase (sdhC) claim 1 , and malate dehydrogenase (mdh).4. The composition of claim 1 , wherein the gene disrupting mutations are in frame gene disrupting mutations.5Edwardsiella ictaluri. A mutant bacterial strain of comprising gene disrupting mutations in genes coding for three proteins in the group consisting of glycine cleavage system (gcvP) claim 1 , succinate ...

POLYNUCLEOTIDES AND VECTORS FOR THE EXPRESSION OF TRANSGENES

Polynucleotides and vectors can be used for the expression of a transgene in cells, such as liver cells. The expression of the transgene from the polynucleotides and vectors can be useful in gene therapy. Various methods can be used for expressing the transgene from the polynucleotides and vectors in liver cells. 1. A polynucleotide , comprising , from 5′ to 3′ , a human ornithine transcarbamylase (hOTC) enhancer , a liver-specific promoter and a transgene , wherein the hOTC enhancer is operably linked to the liver-specific promoter and the liver-specific promoter is operably linked to the transgene.2. The polynucleotide of claim 1 , further comprising an intron between the liver-specific promoter and the transgene.3. The polynucleotide of claim 2 , wherein the intron is a SV40 intron or a beta-globin intron.4. The polynucleotide of any one of - claim 2 , further comprising a polyadenylation signal sequence 3′ of the transgene.5. The polynucleotide of claim 4 , wherein the polyadenylation signal sequence is a BGH-poly(A) signal.6. The polynucleotide of any one of - claim 4 , further comprising a Kozac sequence between the liver-specific promoter and the transgene.7. The polynucleotide of any one of - claim 4 , comprising claim 4 , from 5′ to 3′ claim 4 , a hOTC enhancer claim 4 , a liver-specific promoter claim 4 , an intron claim 4 , a Kozac sequence claim 4 , a transgene claim 4 , and a polyadenylation signal.8. The polynucleotide of any one of - claim 4 , comprising two or more hOTC enhancers operably linked to the liver-specific promoter.9. The polynucleotide of any one of - claim 4 , wherein the liver-specific promoter is a hOTC promoter or a human alpha 1-antitrypsin (hAAT) promoter.10. The polynucleotide of claim 9 , comprising claim 9 , from 5′ to 3′ claim 9 , a hOTC enhancer claim 9 , a hOTC promoter claim 9 , a SV40 intron claim 9 , a Kozac sequence claim 9 , a transgene claim 9 , and a BGH-poly(A) signal.11. The polynucleotide of claim 9 , comprising ...

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A composition comprising a nucleic acid sequence encoding human ornithine transcarbamylase (hOTCase) and expression control sequences which direct expression of hOTC in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over the mature sequence or full length hOTC of SEQ ID NO:1 , and expresses a functional hOTCase , wherein said hOTC nucleic acid sequence is selected from the nucleic acid sequence comprising SEQ ID NO: 5 or a nucleic acid sequence at least about 96 to about 99% identical thereto or a nucleic acid sequence selected from SEQ ID NO: 9 , or a nucleic acid sequence at least about 96 to about 99% identical thereto.2. The recombinant viral vector according to claim 1 , wherein the hOTC nucleic acid sequence has the sequence of SEQ ID NO: 4.3. The recombinant viral vector according to claim 1 , wherein the hOTC nucleic acid sequence has the sequence of SEQ ID NO: 3.4. The recombinant viral vector according to claim 1 , wherein the hOTC nucleic acid sequence has the sequence of SEQ ID NO: 8.5. The recombinant viral vector according to claim 1 , wherein the hOTC is a chimeric OTC comprises a heterologous transit sequence substituted for the native transit sequence of SEQ ID NO: 5 or 9.6. The recombinant viral vector according to claim 1 , wherein the viral vector is selected from an adeno-associated virus (AAV) vector claim 1 , an adenoviral vector claim 1 , and a lentiviral vector.7. The recombinant viral vector according to claim 1 , wherein the expression control sequences further comprise a liver-specific promoter.8. The ...

NOVEL GENETICALLY ENGINEERED MICROORGANISM CAPABLE OF GROWING ON FORMATE, METHANOL, METHANE OR CO2

The present invention relates to a genetically engineered microorganism expressing (i) formate tetrahydrofolate (THF) ligase, methenyi-THF cyclohydrolase and methylene-THF dehydrogenase, (ii) the enzymes of the glycine cleavage system (GCS), (iii) serine deaminase and serine hydroxymethyltransferase (SHMT), (iv) an enzyme increasing the availability of NADPH, and (v) optionally formate dehydrogenase (FDH), and wherein the genetically engineered microorganism has been genetically engineered to express at least one of the enzymes of (i) to (v), wheren said enzyme is not expressed by the corresponding microorganism that has been used to prepare the genetically engineered microorganism, and wherein the enzymes of (i) to (v) are genomically expressed. 1. A genetically engineered microorganism expressing(i) formate tetrahydrofolate (THF) ligase, methenyl-THF cyclohydrolase and methylene-THF dehydrogenase,(ii) the enzymes of the glycine cleavage system (GCS),(iii) serine deaminase and serine hydroxymethyltransferase (SHMT),(iv) an enzyme increasing the availability of NADPH, and(v) optionally formate dehydrogenase (FDH), andwherein the genetically engineered microorganism has been genetically engineered to express at least one of the enzymes of (i) to (v), wheren said enzyme is not expressed by the corresponding microorganism that has been used to prepare the genetically engineered microorganism, andwherein the enzymes of (i) to (v) are genomically expressed.2. The genetically engineered microorganism of claim 1 , wherein the enzymes of (i) to (v) are expressed from genomic safe spots.3. The genetically engineered microorganism of or claim 1 , wherein one or more of the enzymes of (i) to (v) are expressed under the control of a strong constitutive promoter and/or a modified ribosome binding site.4. The genetically engineered microorganism of any one of to claim 1 , wherein the enzyme of (iv) is at least 2-fold claim 1 , preferably at-least 3-fold claim 1 , more preferably ...

Production of odd chain fatty acid derivatives in recombinant microbial cells

Recombinant microbial cells are provided which have been engineered to produce fatty acid derivatives having linear chains containing an odd number of carbon atoms by the fatty acid biosynthetic pathway. Also provided are methods of making odd chain fatty acid derivatives using the recombinant microbial cells, and compositions comprising odd chain fatty acid derivatives produced by such methods.

Microorganisms for the production of putrescine or ornithine and procedure for the production of putrescine or ornithine using the same

<p>SE DESVELA UN MICROORGANISMO MODIFICADO QUE PRODUCE PUTRESCINA U ORNITINA, Y UN PROCEDIMIENTO PARA LA PRODUCCIÓN DE PUTRESCINA U ORNITINA USANDO EL MISMO.</p> <p> A MODIFIED MICROORGANISM THAT PRODUCES PUTRESCINA OR ORNITINE, AND A PROCEDURE FOR THE PRODUCTION OF PUTRESCINA OR ORNITINE USING THE SAME IS DEVELOPED. </p>

Microorganism having inhanced activity of 3-methyl-2-oxobutanoate hydroxymethyltransferase and uses thereof

3-메틸-2-옥소뷰타노에이트 하이드록시 메틸트랜스퍼라아제 변이체, 3-메틸-2-옥소뷰타노에이트 하이드록시 메틸트랜스퍼라아제 활성이 강화된 미생물, 상기 미생물을 포함하는 판토텐산 및/또는 판토산 생산용 조성물, 및 상기 미생물을 배양하는 단계를 포함하는 판토텐산 및/또는 판토산 제조 방법이 제공된다.

Yeast strains engineered to produce ethanol from glycerol

The present invention relates to genetically modified yeast cells comprising exogenous genes coding for pyruvate formate lyase and acetaldehyde dehydrogenase activities. The yeast cells further comprises genetic modifications that improve glycerol utilization such as modifications that increases NAD + -linked glycerol dehydrogenase activity, and preferably one or more of dihydroxyacetone kinase activity and transport of glycerol into the cell. The yeast cell further preferably comprises a functional exogenous xylose isomerase gene and/or functional exogenous genes which confer to the cell the ability to convert L-arabinose into D-xylulose 5-phosphate and they may comprise a genetic modification that increase acetyl-CoA synthetase activity. The process is further characterised in that glycerol is present in or fed into the culture medium, whereby the modified yeast cell ferments the hexoses, pentoses,acetic acid and glycerol to ethanol. The invention further relates to processes for producing a fermentation product and formate from carbon sources comprising glycerol and at least one of a hexose and a pentose, wherein the genetically modified yeast cells are used to ferment the carbon sources to the fermentation product and formate. A preferred fermentation product is ethanol.

Recombinant microorganism producing itaconic acid and production method of itaconic acid using the same

The present invention relates to a recombinant microorganism for itaconic acid. More particularly, the present invention relates to a recombinant microorganism for producing itaconic acid, with improved productivity of itaconic acid through regulation of gene expression involved in acetic acid metabolism, and a production method of itaconic acid using the same. The recombinant microorganism for producing itaconic acid according to the present invention can enhance economic feasibility of itaconic acid by remarkably increasing the production amount of itaconic acid by regulating gene expression involved in acetic acid metabolism, and thus can be used in various fields such as food additives, latex, and synthetic resins where itaconic acid is used.

System of selection and recombinant microorganism and application thereof

微生物存活所需的维生素或其它必需营养素的生物合成途径中的一种或多种基因可被用作有效的选择标记，以鉴定用外源核酸转化的细胞。所述微生物天然不含有或不表达所述一种或多种基因。这使得可进行遗传操作。其允许进行低成本的发酵。其允许生产用于随后的商业用途的必需营养素。

Arginine supplementation to improve efficiency in gas fermenting acetogens

The invention provides methods for improving efficiency of fermentation by arginine supplementation, and genetically modified bacterium for use therefor. More particularly the invention provides methods for (i) increasing the production ATP intensive products with arginine supplementation, (ii) increasing utilization of arginine by a C1-fixing bacterium; and (iii) providing C1-fixing bacterium with optimized arginine de-aminase pathways.

Corynebacterium glutamicum variety producing l-arginine and method for fabricating the same

An L-arginine producing microorganism is provided to over-express an argF2 gene, thereby producing an L-arginine with high yield and being usefully used for human medicinal and pharmaceutical industries. An L-arginine producing microorganism is characterized in that it is transformed by a polynucleotide encoding an argF(NCBI access no. Ncg10990) polypeptide. In the microorganism, the argF2 polypeptide has an amino acid sequence described as SEQ ID : NO. 1, the polynucleotide is described as SEQ ID : NO. 2 and the microorganism is Corynebacterium sp. and is deposited as a deposition no. KCCM10819P or KCCM10821P. The L-arginine producing microorganism is cultured in order to produce L-arginine.

Pseudomonas aeruginosa PcrV-linked antigen vaccine

Putrescine producing microorganisms, and the putrescine production method by using these microorganisms

FIELD: chemistry.SUBSTANCE: group of inventions relates to the putrescine producing microorganism, and to the use of said microorganism putrescine production method. In the proposed microorganism, the protein activity is increased having the amino acid sequence, as presented in the SEQ ID NO: 21 or 23, compared to the said protein activity in the wild-type microorganism. Putrescine production method includes the said microorganism culturing with the cells culture production, and the putrescine isolation from the cultured microorganism or cells culture.EFFECT: group of inventions provides putrescine production in high yield.10 cl, 11 tbl, 6 ex, 2 dwg РОССИЙСКАЯ ФЕДЕРАЦИЯ ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) (19) RU (11) (13) 2 665 825 C2 (51) МПК C12N 1/21 (2006.01) C12N 15/52 (2006.01) C12N 9/02 (2006.01) C12N 9/10 (2006.01) C12N 9/12 (2006.01) C12N 9/88 (2006.01) C12P 13/00 (2006.01) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/52 (2006.01); C12N 9/0008 (2006.01); C12N 9/1018 (2006.01); C12N 9/1029 (2006.01); C12N 9/1096 (2006.01); C12N 9/1217 (2006.01); C12N 9/88 (2006.01); C12P 13/001 (2006.01); C12Y 102/01038 (2006.01); C12Y 201/03003 (2006.01); C12Y 203/01035 (2006.01); C12Y 206/01011 (2006.01); C12Y 207/02008 (2006.01); C12Y 401/01007 (2006.01) 2015142261, 25.02.2014 (24) Дата начала отсчета срока действия патента: 25.02.2014 04.09.2018 Приоритет(ы): (30) Конвенционный приоритет: 20.03.2013 KR 10-2013-0030020; 14.02.2014 KR 10-2014-0017243 (43) Дата публикации заявки: 26.04.2017 Бюл. № 12 (56) Список документов, цитированных в отчете о поиске: WO 2012077995 A2, 14.06.2012. RU (85) Дата начала рассмотрения заявки PCT на национальной фазе: 20.10.2015 2433180 C2, 10.11.2011. SCHNEIDER J. ET AL. Improving putrescine production by Corynebacterium glutamicum by fine-tuning ornithine transcarbamoylase activity using a plasmid addiction system // Appl Microbiol Biotechnol (2012) 95:169-178. ZAHOOR A. ET AL. Metabolic engineering of ...

Process for the fermentative production of D-pantothenic acid using coryneform bacteria

Die Erfindung betrifft in Mikroorganismen der Gattung Corynebacterium replizierbare, gegebenenfalls rekombinante DNA mit der Herkunft Corynebacterium, die zumindest eine der folgenden Nucleotidsequenzen enthält, ausgewählt aus der Gruppe: DOLLAR A a) codierend für das panB-Gen (Ketopantoathydroxy-methyltransferese), dargestellt in der SEQ-ID-No.1, DOLLAR A b) codierend für das panC-Gen (Pantothenatsynthetase), dargestellt in der SEQ-ID-No. 1, insbesondere das panBC-Operon und gegebenenfalls DOLLAR A c) codierend für das ilvD-Gen (Dihydroxysäuredehydratase), dargestellt durch die SEQ-ID-No.4, DOLLAR A und ein Verfahren zur fermentativen Herstellung von D-Pantothensäure unter Verwendung von Mikroorganismen der Gattung Corynebacterium, in denen die genannten Gene verstärkt werden. In microorganisms of the genus Corynebacterium, the invention relates to replicable, optionally recombinant DNA with the origin Corynebacterium, which contains at least one of the following nucleotide sequences, selected from the group: DOLLAR A a) coding for the panB gene (ketopantoate hydroxy methyl transfer), shown in FIG SEQ-ID-No.1, DOLLAR A b) coding for the panC gene (pantothenate synthetase), shown in SEQ-ID-No. 1, in particular the panBC operon and optionally DOLLAR A c) coding for the ilvD gene (dihydroxy acid dehydratase), represented by SEQ-ID-No.4, DOLLAR A and a process for the fermentative production of D-pantothenic acid using microorganisms the genus Corynebacterium, in which the genes mentioned are amplified.

Process for the microbial production of L-valine

Verfahren zur mikrobiellen Herstellung von L-Valin, bei dem die Dihydroxysäuredehydratase-(ilvD) Aktivität und/oder die ilvD-Genexpression in einem Mikroorganismus verstärkt wird, dadurch gekennzeichnet, dass die Aktivität des Enzyms Ketopanthoathydroxymethyltransferase (pan B) und/oder des Enzyms Pantothenatligase (pan C) und/oder Aspartatdecarboxylase abgeschwächt oder ausgeschaltet ist. method for the microbial production of L-valine, in which the dihydroxy acid dehydratase (ilvD) activity and / or amplifying ilvD gene expression in a microorganism, characterized in that the activity of the enzyme Ketopanthoathydroxymethyltransferase (pan B) and / or the enzyme pantothenate ligase (pan C) and / or aspartate decarboxylase attenuated or is off.

Methods and microorganisms for production of panto-compounds

Dna encoding novel enzyme having d-serine synthase activity, method of producing the enzyme and method of producing d-serine by using the same

본 발명은, 글리신과 포름알데히드로부터 D-세린을 합성하는 활성을 갖는 신규한 효소를 코드하는 DNA, 상기 DNA를 벡터에 삽입하여 이루어지는 재조합 DNA, 상기 재조합 DNA로 형질전환한 형질전환체, 및 상기 효소를 이용한 포름알데히드와 글리신으로부터 D-세린을 제조하는 방법에 관한 것이다.

Microorganisms for producing ornithine and method of producing ornithine using said microorganisms

FIELD: biotechnology. SUBSTANCE: present invention relates to biotechnology and represents a modified microorganism of the genus Corynebacterium , which produces ornithine, where the activity of N-acetylglutamate synthase from E. coli and acetylornithine deacetylase from E. coli are introduced into said microorganism of the genus Corynebacterium , having acetylglutamate synthase or ornithine acetyltransferase activity (ArgJ). EFFECT: invention increases ornithine production. 12 cl, 2 dwg, 11 tbl, 7 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 721 852 C1 (51) МПК C12N 15/77 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/52 (2020.02); C12N 9/1029 (2020.02); C12N 9/80 (2020.02); C12N 9/88 (2020.02); C12P 13/001 (2020.02) (21)(22) Заявка: 2019126098, 19.07.2016 19.07.2016 Дата регистрации: 25.05.2020 20.07.2015 KR 10-2015-0102624 Номер и дата приоритета первоначальной заявки, из которой данная заявка выделена: 2018104512 20.07.2015 Адрес для переписки: 191036, Санкт-Петербург, а/я 24, "НЕВИНПАТ" (54) Микроорганизмы для получения орнитина и способ получения орнитина с использованием указанных микроорганизмов (57) Реферат: Настоящее изобретение относится к указанный микроорганизм рода Corynebacterium, биотехнологии и представляет собой обладающий активностью модифицированный микроорганизм рода ацетилглутаматсинтазы или Corynebacterium, продуцирующий орнитин, где орнитинацетилтрансферазы (ArgJ). Изобретение активности N-ацетилглутаматсинтазы из E. coli и позволяет повысить продукции орнитина. 2 н. и ацетилорнитиндезацетилазы из E. coli введены в 10 з.п. ф-лы, 2 ил., 11 табл, 7 пр. R U 2 7 2 1 8 5 2 (56) Список документов, цитированных в отчете о поиске: KR 101053429 B1, 03.08.2011. DE 1642678 A1, 03.06.1971. CN 102191291 A, 21.09.2011. RU 2433180 C2, 10.11.2011. RU 2316588 C1, 10.02.2008. Стр.: 1 C 1 C 1 (45) Опубликовано: 25.05.2020 Бюл. № 15 (73) Патентообладатель(и): СИДЖЕЙ ЧЕИЛДЖЕДАНГ КОРПОРЕЙШН (KR) 2 ...

Microorganisms for producing putrescine or ornithine and method of producing putrescine or ornithine using said microorganisms

Группа изобретений относится к модифицированному микроорганизму рода Corynebacterium, продуцирующему путресцин, и способу получения путресцина с использованием указанного микроорганизма. Предложен модифицированный микроорганизм рода Corynebacterium, в котором активности N-ацетилглутаматсинтазы из E. coli и ацетилорнитиндезацетилазы из E. coli введены в указанный микроорганизм рода Corynebacterium, обладающий активностью ацетилглутаматсинтазы или орнитинацетилтрансферазы (ArgJ). Также предложен способ получения путресцина, включающий культивирование указанного модифицированного микроорганизма рода Corynebacterium в среде и выделение путресцина из культивированного микроорганизма или среды. Группа изобретений позволяет получить путресцин с увеличенным выходом. 2 н. и 15 з.п. ф-лы, 2 ил., 11 табл., 7 пр. РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 708 165 C2 (51) МПК C12N 15/77 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/52 (2019.08); C12N 9/1029 (2019.08); C12N 9/1217 (2019.08); C12N 9/80 (2019.08); C12N 9/88 (2019.08); C12N 9/93 (2019.08); C12P 13/001 (2019.08); C12P 13/10 (2019.08); C12Y 203/01001 (2019.08); C12Y 203/01008 (2019.08); C12Y 203/01035 (2019.08); C12Y 207/02001 (2019.08); C12Y 305/01016 (2019.08); C12Y 401/01017 (2019.08); C12Y 602/01001 (2019.08) 2018104512, 19.07.2016 (24) Дата начала отсчета срока действия патента: 19.07.2016 04.12.2019 Приоритет(ы): (30) Конвенционный приоритет: 20.07.2015 KR 10-2015-0102624 (43) Дата публикации заявки: 20.08.2019 Бюл. № 23 (56) Список документов, цитированных в отчете о поиске: US 8497098 D2, 30.07.2013. DOU W. et al. Improvement of L-arginine Production by Overexpression of a Bifunctional Ornithine Acetyltransferase in Corynebacterium Crenatum. Applied Biochemistry and Biotechnology, 2011, vol. 165, pages 845-855. база данных GenBank:EZG54138.1, 09.04.2014. база данных GenBank:BAE77354.1, 20.11.2008. RU 2550269 C2, (см. прод.) (85) Дата начала ...

Microorganisms for obtaining putrescine and method for obtaining putrescine with application thereof

FIELD: chemistry. SUBSTANCE: invention relates to biotechnology, namely to putrescine-producing microorganism and method for obtaining putrescine with thereof application. Putrescine-producing microorganism is modified in such a way that activity of ornithinecarbamoyltransferase and protein, participating in glutamate (NCgl1221) export in comparison with their endogenic activities is deleted in it, and ornithinedecarboxilase (ODC) activity is introduced into microorganism. EFFECT: invention makes it possible to obtain putrescine with high degree of effectiveness. 13 cl, 3 dwg, 8 tbl, 5 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (51) МПК C12N 1/21 (13) 2 573 923 C2 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ (21)(22) Заявка: ИЗОБРЕТЕНИЯ К ПАТЕНТУ 2013131033/10, 08.12.2011 (24) Дата начала отсчета срока действия патента: 08.12.2011 Приоритет(ы): (30) Конвенционный приоритет: R U 08.12.2010 KR 10-2010-0124867; 07.12.2011 KR 10-2011-0130595 (72) Автор(ы): ЧОИ Хианг (KR), ЛИ Киоунг Мин (KR), КАНГ Мин Сун (KR), ДЗХОН Сунг Хоо (KR), УМ Хие Вон (KR), ЧОЙ Су Дзин (KR), ЛИ Хан Вон (KR), ШИН Соо Ан (KR) (43) Дата публикации заявки: 20.01.2015 Бюл. № 2 C 2 C 2 (56) Список документов, цитированных в отчете о поиске: SCHNEIDER J et.al. Putrescine production by engineered Corynebacterium glutamicum, Appl Microbiol Biotechnol. 2010 Oct;88(4):859-68. doi: 10.1007/s00253-010-2778-x. Epub 2010 Jul 27. HASHIMOTO K. et.al. The protein encoded by NCgl1221 in Corynebacterium glutamicum functions as a mechanosensitive channel, Biosci Biotechnol Biochem. (см. прод.) (73) Патентообладатель(и): СиДжей ЧЕИЛДЗЕДАНГ КОРПОРЕЙШН (KR) (85) Дата начала рассмотрения заявки PCT на национальной фазе: 08.07.2013 2 5 7 3 9 2 3 (86) Заявка PCT: R U 2 5 7 3 9 2 3 (45) Опубликовано: 27.01.2016 Бюл. № 3 KR 2011/009478 (08.12.2011) (87) Публикация заявки PCT: WO 2012/077995 (14.06.2012) Адрес для переписки: 129090, Москва, ул. Б. Спасская, 25, строение 3, ООО "Юридическая фирма ...

Methods and compositions of otc constructs and vectors

Provided herein are methods and compositions related to nucleic acids encoding ornithine transcarbamylase (OTC), such as nucleic acids comprising an OTC codon- optimized sequence, as well as related vectors, such as AAV vectors. Also, provided are methods for administering AAV vectors that comprise a sequence that encodes an enzyme associated with an urea cycle disorder and an expression control sequence, in combination with synthetic nanocarriers coupled to an immunosuppressant.

Microorganism which produces putrescine or ornithine, and a method of producing putrescine or ornithine using said microorganism

FIELD: biotechnology.SUBSTANCE: disclosed is a modified microorganism of the genus Corynebacterium producing putrescine with reduced activity of the sugar metabolism transcription (SugR) regulator compared to its endogenous activity and high activity of citrate synthase (GltA) compared to its endogenous activity. Disclosed is a method of producing putrescine using said microorganism.EFFECT: group of inventions makes it possible to increase productivity of the specified modified microorganism by putrescine in comparison with non-modified microorganism.15 cl, 7 tbl, 8 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 691 303 C1 (51) МПК C12N 15/77 (2006.01) C12N 9/10 (2006.01) C12P 13/00 (2006.01) C12P 13/10 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/77 (2018.08); C12N 9/1025 (2018.08); C12P 13/001 (2018.08); C12P 13/10 (2018.08); C12Y 203/03001 (2018.08) (21)(22) Заявка: 2017145909, 29.03.2016 29.03.2016 Дата регистрации: 11.06.2019 24.06.2015 KR 10-2015-0090021 (45) Опубликовано: 11.06.2019 Бюл. № 17 (73) Патентообладатель(и): СИДЖЕЙ ЧЕИЛДЖЕДАНГ КОРПОРЕЙШН (KR) (85) Дата начала рассмотрения заявки PCT на национальной фазе: 24.01.2018 (56) Список документов, цитированных в отчете о поиске: JP 2009254323 A, 05.11.2009. KR R U 2 6 9 1 3 0 3 C 1 KR 2016/003198 (29.03.2016) (87) Публикация заявки PCT: WO 2016/208854 (29.12.2016) Адрес для переписки: 191036, Санкт-Петербург, а/я 24, "НЕВИНПАТ" (54) Микроорганизм, продуцирующий путресцин или орнитин, и способ получения путресцина или орнитина с использованием этого микроорганизма (57) Реферат: Группа изобретений относится к области с ее эндогенной активностью. Предложен способ биотехнологии. Предложен модифицированный получения путресцина с использованием микроорганизм рода Corynebacterium, указанного микроорганизма. Группа изобретений продуцирующий путресцин, с пониженной позволяет увеличить продуктивность указанного активностью регулятора транскрипции ...

Inducible COMT_II promoter, chimeric gene containing same and plants transformed therewith

The invention concerns a novel regulating COMTII promoter sequence inducible in response to a mechanical or chemical injury, or in response to aggression by a pathogenic agent, in particular bacterial, fungal or viral, or by an insect or a threadworm. The invention also concerns a chimera gene (or expression cassette) comprising the inventive regulating promoter sequence controlling the expression of a heterologous coding sequence and a host organism comprising said chimera gene, transformed plants containing it and the seeds of said transformed plants.

Putrescine or ornithine producing microorganism and method for producing putrescine or ornithine using the same

本発明はプトレシンまたはオルニチンを生産するための組み換え微生物及びそれを用いてプトレシンまたはオルニチンを生産する方法に関する。 【選択図】図２

Novel formate-dependent phosphoribosylglycinamide formyltransferase variant and a method for producing IMP using the same

The present application relates to a novel formate-dependent phosphoribosylglycinamide formyltransferase variant, a Corynebacterium stationis strain comprising the variant, and a method for producing IMP using the strain. According to the present invention, high yield IMP production is possible.

Microorganism producing putrescine or ornithine and method for producing putrescine or ornithine using this microorganism

FIELD: microbiology.SUBSTANCE: group of inventions relates to a modified microorganism for producing ornithine and to a method for producing ornithine using this microorganism. A modified microorganismCorynebacterium glutamicumis proposed, producing ornithine with reduced activity of a regulator of sugar metabolism transcription (SugR) in comparison with endogen activity ofCorynebacterium glutamicumand increased activity of citrate synthase (GltA) in comparison with endogen activity ofCorynebacterium glutamicum, where thisCorynebacterium glutamicumhas a capability of producing ornithine. A method for producing ornithine using the specified microorganism is also proposed.EFFECT: group of inventions provides producing ornithine with high yield.8 cl, 7 tbl, 8 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 759 956 C1 (51) МПК C12N 15/77 (2006.01) C12N 9/10 (2006.01) C12N 9/88 (2006.01) C12P 13/10 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/77 (2021.08); C12N 9/1018 (2021.08); C12N 9/1025 (2021.08); C12N 9/88 (2021.08); C12P 13/10 (2021.08); C12Y 201/03003 (2021.08); C12Y 203/03001 (2021.08); C12Y 401/01017 (2021.08) (21)(22) Заявка: 2019103640, 29.03.2016 29.03.2016 Дата регистрации: 19.11.2021 24.06.2015 KR 10-2015-0090021 Номер и дата приоритета первоначальной заявки, из которой данная заявка выделена: 2017145909 24.06.2015 C 1 2 7 5 9 9 5 6 Адрес для переписки: 191036, Санкт-Петербург, а/я 24, "НЕВИНПАТ" (54) Микроорганизм, продуцирующий путресцин или орнитин, и способ получения путресцина или орнитина с использованием этого микроорганизма (57) Реферат: Группа изобретений относится к glutamicum и повышенной активностью модифицированному микроорганизму для цитратсинтазы (GltA) по сравнению с эндогенной получения орнитина и к способу получения активностью Corynebacterium glutamicum, где этот орнитина с использованием этого Corynebacterium glutamicum обладает микроорганизма. Предложен модифицированный ...

Mutant microorganism with improved productivity of branched amino acid and method for preparing it using the same

A mutant microorganism obtained by mutation and metabolic flux operation is provided to produce a branched amino acid, particularly L-valine with high efficiency, thereby being industrially useful for generating the L-valine. A method for preparing a mutant microorganism with high L-valine productivity is characterized in that a gene encoding an enzyme involving with L-isoleucine bio-synthesis such as ilvA(a gene encoding threonine dehydratase), a gene encoding an enzyme involving with L-leucine bio-synthesis such as leuA(a gene encoding 2-isopropylmalate synthase), and a gene encoding an enzyme involving with D-pantothenic acid bio-synthesis such as panB(a gene encoding 3-methyl-2-oxobutanoate hydroxymethyl transferase) are attenuated or deleted and a gene encoding an enzyme involving with L-valine bio-synthesis is mutated by deleting lacI(a gene encoding lac operon repressor), removing feedback inhibition of ilvH(acetohydroxy acid synthase isozyme III) gene, substituting a native promoter including an attenuator of ilvGMEDA(acetohydroxy acid synthase isozyme I) and ilvBN(acetohydroxy acid synthase isozyme II) with a strong promoter, or introducing an expression vector including a strong promoter. A method for preparing L-valine comprises the steps of: (a) culturing the mutant microorganism having high L-valine productivity; and (b) recovering the L-valine from a culture solution of the microorganism.

MICROORGANISMS WITH THE IMPROVED ORNITINE-PRODUCING ABILITY, AND METHOD OF OBTAINING ORNITINE USING THEM

1. Микроорганизм, имеющий улучшенную орнитин-продуцирующую способность, где активности орнитинкарбамоилтрансферазы и белка, участвующего в экспорте глутамата (NCgl1221), модифицированы таким образом, что они являются аттенуированными в сравнении с их эндогенными активностями.2. Микроорганизм по п. 1, где орнитинкарбамоилтрансфераза имеет аминокислотную последовательность SEQ ID NO:18 или аминокислотную последовательность, имеющую 70% или более высокую гомологию с этой последовательностью.3. Микроорганизм по п. 1, где белок, участвующий в экспорте глутамата, имеет аминокислотную последовательность SEQ ID NO:20 или аминокислотную последовательность, имеющую 70% или более высокую гомологию с этой последовательностью.4. Микроорганизм по п. 1, где активность орнитинкарбамоилтрансферазы и белка, участвующего в экспорте глутамата, является аттенуированной способом, выбранным из группы, состоящей из (1) частичной или полной делеции гена, кодирующего этот белок, (2) модификации регуляторной последовательности экспрессии для супрессии экспрессии гена, (3) модификации последовательности гена на хромосоме для уменьшения активности этого белка и (4) их комбинации.5. Микроорганизм по п. 1, где активности ацетил-гамма-глутамилфосфатредуктазы (ArgC), ацетилглутаматсинтазы или орнитинацетилтрансферазы (ArgJ), ацетилглутаматкиназы (ArgB) и ацетилорнитинаминотрансферазы (ArgD) являются дополнительно усиленными в сравнении с их эндогенными активностями.6. Микроорганизм по п. 5, где каждый из ArgC, ArgJ, ArgB и ArgD имеет аминокислотную последовательность SEQ ID NO:23, 25, 27 и 29 или аминокислотную последовательность, имеющую 70% или более высокую гомологию с этой послед� РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (51) МПК C12N 1/21 (13) 2013 131 039 A (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2013131039/10, 08.12.2011 (71) Заявитель(и): СиДжей ЧЕИЛДЗЕДАНГ КОРПОРЕЙШН (KR) Приоритет(ы): (30) Конвенционный приоритет: 08.12.2010 KR ...

Bacillus subtilis and construction method and application thereof

本发明属于微生物领域，公开了一种枯草芽孢杆菌及其构建方法与应用。本发明所述枯草芽孢杆菌其为枯草芽孢杆菌菌株中purD P116L 、purR A65D 或guaB G279R 中至少一个位点发生了点突变。purD P116L 、purR A65D 或guaB G279R 单一位点突变分别获得枯草芽孢杆菌B.subtilis A1、A2、A3菌株。三个单一点突变质粒依次转化B.subtilis168(Δupp)菌株获得三个位点均发生点突变的枯草芽孢杆菌B.subtilis A5菌株。本发明所述枯草芽孢杆菌为腺苷高产菌株，能有效积累腺苷，提高腺苷的产量，为腺苷的工业化生产奠定了基础。

Microorganism having inhanced activity of 3-methyl-2-oxobutanoate hydroxymethyltransferase and uses thereof

3-메틸-2-옥소뷰타노에이트 하이드록시 메틸트랜스퍼라아제 변이체, 3-메틸-2-옥소뷰타노에이트 하이드록시 메틸트랜스퍼라아제 활성이 강화된 미생물, 상기 미생물을 포함하는 판토텐산 및/또는 판토산 생산용 조성물, 및 상기 미생물을 배양하는 단계를 포함하는 판토텐산 및/또는 판토산 제조 방법이 제공된다.

Production of malonyl-CoA derived products via anaerobic pathways

The present invention provides for novel metabolic pathways to convert biomass and other carbohydrate sources to malonyl-CoA derived products, such as hydrocarbons and other bioproducts, under anaerobic conditions and with the net production of ATP. More specifically, the invention provides for a recombinant microorganism comprising one or more native and/or heterologous enzymes that function in one or more engineered metabolic pathways to achieve conversion of a carbohydrate source to, e.g., long-chain hydrocarbons and hydrocarbon derivatives, wherein the one or more native and/or heterologous enzymes is activated, upregulated, downregulated, or deleted. The invention also provides for processes to convert biomass to malonyl-CoA derived products which comprise contacting a carbohydrate source with a recombinant microorganism of the invention.

Pichia ciferrii cells and their use

Gegenstand der Erfindung sind gentechnisch modifizierte Pichia ciferri Zellen, deren Verwendung sowie ein Verfahren zur Herstellung von Sphingoidbasen und Sphingolipiden. The invention relates to genetically modified Pichia ciferri cells, their use and a process for the preparation of sphingoid bases and sphingolipids.

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

본 발명은 퓨트레신 또는 오르니틴을 생산하기 위한 재조합 미생물 및 이를 이용하여 퓨트레신 또는 오르니틴을 생산하는 방법에 관한 것이다. The present invention relates to a recombinant microorganism for producing putrescine or ornithine, and a method for producing putrescine or ornithine using the recombinant microorganism.

- - L-methionine producing microorganism and method of producing L-methionine using the microorganism

본 발명은 L-쓰레오닌을 생산하는 균주에서 L-메씨오닌 생합성에 관여하는 단백질을 과발현시켜 고수율로 L-메씨오닌을 생산하는 균주를 제조하는 방법, 이러한 방법으로 제조된 균주 및 이러한 균주를 이용하여 L-메씨오닌을 생산하는 방법에 관한 것이다. The present invention provides a method for producing L-methionine in high yield by overexpressing a protein involved in L-methionine biosynthesis in a strain producing L-threonine, a strain prepared by such a method and a strain produced by such a method. It relates to a method for producing L-methionine using. 쓰레오닌 디하이드라타제, O-숙시닐호모세린 리아제, 시스타씨오닌 베타 리아제, 5,10-메틸렌테트라하이드로폴레이트 리덕타제, 세린 하이드록시메틸트랜스페라제, L-메씨오닌, S-아데노실-메씨오닌 Threonine dehydratase, O-succinylhomoserine lyase, cystathionine beta lyase, 5,10-methylenetetrahydrofolate reductase, serine hydroxymethyltransferase, L-methionine, S-adeno Syl-methionine

Patent RU2018104512A3

РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2018 104 512 A (51) МПК C12N 15/77 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2018104512, 19.07.2016 (71) Заявитель(и): СИДЖЕЙ ЧЕИЛДЖЕДАНГ КОРПОРЕЙШН (KR) Приоритет(ы): (30) Конвенционный приоритет: 20.07.2015 KR 10-2015-0102624 (85) Дата начала рассмотрения заявки PCT на национальной фазе: 20.02.2018 (86) Заявка PCT: (87) Публикация заявки PCT: WO 2017/014532 (26.01.2017) R U (54) Микроорганизмы для получения путресцина или орнитина и способ получения путресцина или орнитина с использованием указанных микроорганизмов (57) Формула изобретения 1. Модифицированный микроорганизм рода Corynebacterium, продуцирующий путресцин или орнитин, в который введены активности N-ацетилглутаматсинтазы из Е. coli и ацетилорнитиндезацетилазы из Е. coli. 2. Модифицированный микроорганизм по п. 1, где N-ацетилглутаматсинтаза из Е. coli состоит из аминокислотной последовательности SEQ ID NO: 1. 3. Модифицированный микроорганизм по п. 1, где ацетилорнитиндезацетилаза из Е. coli состоит из аминокислотной последовательности SEQ ID NO: 3. 4. Модифицированный микроорганизм по п. 1, где микроорганизм рода Corynebacterium представляет собой Corynebacterium glutamicum. 5. Модифицированный микроорганизм по п. 1, где активность фосфотрансацетилазного и ацетаткиназного оперона (оперона pta-ackA) дополнительно усилена по сравнению с его эндогенной активностью. 6. Модифицированный микроорганизм по п. 5, где фосфотрансацетилазный и ацетаткиназный оперон состоит из аминокислотной последовательности SEQ ID NO: 5 или SEQ ID NO: 7. 7. Модифицированный микроорганизм по п. 1, в который дополнительно введена активность ацетил-КоА-синтетазы (acs) из Е. coli. 8. Модифицированный микроорганизм по п. 7, где ацетил-КоА-синтетаза состоит из аминокислотной последовательности SEQ ID NO: 9. Стр.: 1 A 2 0 1 8 1 0 4 5 1 2 A Адрес для переписки: 191036, Санкт-Петербург, а/я 24, "НЕВИНПАТ" 2 0 1 8 1 0 4 5 1 2 KR 2016/ ...

Pichia ciferrii cells and use thereof

本发明涉及遗传学修饰的毕赤酵母细胞及其应用，还涉及生产鞘氨醇碱和鞘脂的方法。

Микроорганизмы, имеющие улучшенную орнитин-продуцирующую способность, и способ получения орнитина с их использованием

Изобретение относится к биотехнологии и представляет собой микроорганизм, имеющий орнитин-продуцирующую способность, где активности орнитинкарбамоилтрансферазы и белка, участвующего в экспорте глутамата (NCgl1221), модифицированы таким образом, что они являются делетированными в сравнении с их эндогенными активностями. Изобретение относится также к способу получения орнитина с использованием этого микроорганизма. Изобретение позволяет получать L-орнитин с высокой степенью эффективности. 2 н. и 9 з.п. ф-лы, 3 ил., 6 табл., 4 пр. РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 567 669 C2 (51) МПК C12N 1/21 (2006.01) C12P 13/04 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ (21)(22) Заявка: ИЗОБРЕТЕНИЯ К ПАТЕНТУ 2013131039/10, 08.12.2011 (24) Дата начала отсчета срока действия патента: 08.12.2011 Приоритет(ы): (30) Конвенционный приоритет: R U 08.12.2010 KR 10-2010-0124866; 07.12.2011 KR 10-2011-0130594 (72) Автор(ы): ЧОИ Хианг (KR), ЛИ Киоунг Мин (KR), КАНГ Мин Сун (KR), ДЗХОН Сунг Хоо (KR), УМ Хие Вон (KR), ЧОЙ Су Дзин (KR), ЛИ Хан Вон (KR), ШИН Соо Ан (KR) (43) Дата публикации заявки: 20.01.2015 Бюл. № 2 C 2 C 2 (56) Список документов, цитированных в отчете о поиске: SCHNEIDER J et.al. Production of the amino acids l-glutamate, l-lysine, l-ornithine and l-arginine from arabinose by recombinant Corynebacterium glutamicum, J Biotechnol. 2011 Jul 10;154(2-3):191-8. doi: 10.1016/ j.jbiotec.2010.07.009. Epub 2010 Jul 16.RU 2316588 C1, 10.02.2008.HASHIMOTO K. et.al. The protein encoded by NCgl1221 in Corynebacterium (см. прод.) (73) Патентообладатель(и): СиДжей ЧЕИЛДЗЕДАНГ КОРПОРЕЙШН (KR) (85) Дата начала рассмотрения заявки PCT на национальной фазе: 08.07.2013 2 5 6 7 6 6 9 (86) Заявка PCT: R U 2 5 6 7 6 6 9 (45) Опубликовано: 10.11.2015 Бюл. № 31 KR 2011/009477 (08.12.2011) (87) Публикация заявки PCT: WO 2012/077994 (14.06.2012) Адрес для переписки: 129090, Москва, ул. Б. Спасская, 25, строение 3, ООО "Юридическая фирма Городисский и Партнеры" (54 ...

Putrescine-producing microorganisms and a method for producing putrescine using these microorganisms

РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2015 142 261 A (51) МПК C12N 1/21 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2015142261, 25.02.2014 (71) Заявитель(и): СиДжей ЧеилДжеданг Корпорейшн (KR) Приоритет(ы): (30) Конвенционный приоритет: 20.03.2013 KR 10-2013-0030020; 14.02.2014 KR 10-2014-0017243 (85) Дата начала рассмотрения заявки PCT на национальной фазе: 20.10.2015 KR 2014/001509 (25.02.2014) (87) Публикация заявки PCT: WO 2014/148743 (25.09.2014) R U (54) Микроорганизмы, продуцирующие путресцин, и способ получения путресцина с использованием этих микроорганизмов (57) Формула изобретения 1. Микроорганизм, продуцирующий путресцин, где указанный микроорганизм модифицирован для усиления активности белка, имеющего аминокислотную последовательность, представленную в SEQ ID NO: 21 или 23. 2. Микроорганизм, продуцирующий путресцин, по п. 1, дополнительно модифицированный для ослабления активности орнитинкарбамоилтрансферазы (ArgF) и белка (NCgl1221), вовлеченного в экспорт глутамата, по сравнению с эндогенной активностью и для усиления активности орнитиндекарбоксилазы (ODC). 3. Микроорганизм, продуцирующий путресцин, по п. 2, где орнитинкарбамоилтрансфераза (ArgF) имеет аминокислотную последовательность, представленную в SEQ ID NO: 29, белок (NCgl1221) вовлеченный в экспорт глутамата, имеет аминокислотную последовательность, представленную в SEQ ID NO: 30, и орнитиндекарбоксилаза (ODC) имеет аминокислотную последовательность, представленную в SEQ ID NO: 33. 4. Микроорганизм, продуцирующий путресцин, по п. 1, дополнительно модифицированный для усиления активности ацетил-гамма-глутамилфосфатредуктазы (ArgC), ацетилглутаматсинтазы или орнитинацетилтрансферазы (ArgJ), ацетилглутаматкиназы (ArgB) и ацетилорнитинаминотрансферазы (ArgD) по сравнению с эндогенной активностью. 5. Микроорганизм, продуцирующий путресцин, по п. 4, где ацетил-гаммаСтр.: 1 A 2 0 1 5 1 4 2 2 6 1 A Адрес для переписки: 191036, Санкт- ...

mRNA for use in treatment of human genetic diseases

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification conferring stability to the RNA, as well as related methods, are disclosed.

Ornithine carbamoyl transferase sequence and uses thereof

The invention provides ornithine carbamoyltransferase polypeptides and DNA (RNA) encoding ornithine carbamoyltransferase polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing ornithine carbamoyltransferase polypeptides to screen for antibacterial compounds.