Biomarker for prediction of sensitive skin and test kit using the same

In the present invention refers to sensitive skin or in the skin tissue derived from compared to former insensitive with a selective ratio relative to the expression of gene expression and 19 is reduced was buys gene species 35. Therefore, by a measure of the degree to of expression said gene, it is judged whether the vehicle is sensitive skin or in the is enabled.

1 table a in sensitive skin determined expression compared to former from the skin of a subject with a selective ratio relative to the precursor and CaO precursor a gene, a table 2 revealed a gene expressed is reduced. With reference to, in table a the NCBI genebank accession ID of GBAcc of. mixture by the addition of an initiator.

One embodiment: an sensitive skin or in the markers are diagnostic biochip, said table 1 of a gene, HLA-C, TCEA1, IGHA1, TFRC, S100A8, LOC652128, SERPINB13, EHF, FLJ10781, TP63, CDH1, TNFRSF19, GM2A, FKBP5, PI3, PSPH, HORMAD1, SPRR2G ACTR2 and one or more selected from the group consisting of includes gene, said gene expression dose sensitive from the skin a elevated relative to former characterized in that. Said gene expression dose compared to former from the skin susceptibility 2 to it is preferable that the.

Other embodiment: an sensitive skin or in the markers are diagnostic biochip, said table NFYA 1 of a gene transcription of the transcription factor gene, ATF4 and MYOG selected from the group consisting of includes at least one gene, said gene expression dose sensitive from the skin a elevated relative to former characterized in that. Said gene expression dose compared to former from the skin susceptibility 2 to it is preferable that the.

Another embodiment: an sensitive skin or in the markers are diagnostic biochip, said table 2 of a gene, KBTBD10, TTN, ACTA1, MYBPC1, MB, MYOZ1, TPM1, CA3, CASQ1, ENO3, RBP4, GYG2, PPP1 R1A, PYGM, G0S2, NEB, H19, LIPE, PLIN, H19, GPAM, FABP4, CIDEC, LPL, PCK1, ADIPOQ, PDE3B, ACVR1C, LPL, FABP4, DKFZP761N09121, TNMD, CRISP3 and ATP6V1B1, C8orf22 selected from the group consisting of includes at least one gene, said gene expression dose sensitive from the skin is reduced relative to former is characterised in that it has a. Said gene expression dose susceptibility compared to former from the skin it is preferable that the reduced to 50% hereinafter.

Another embodiment: an sensitive skin or in the markers are diagnostic biochip, said table 2 of a gene transcription of the transcription factor gene NFKB1, SRF, RELA, TCF3, POU2F1, JUN and USF selected from the group consisting of includes at least one gene, said gene expression dose sensitive from the skin is reduced relative to former is characterised in that it has a. Said gene expression dose susceptibility compared to former from the skin it is preferable that the reduced to 50% hereinafter.

Furthermore, the present invention refers to said table 1 or 2 gene of protein and the uses coded from sensitive skin diagnosis biomarker for provides.

One embodiment: an sensitive skin or in the markers are diagnostic biochip, HLA-C, TCEA1, IGHA1, TFRC, S100A8, LOC652128, SERPINB13, EHF, FLJ10781, TP63, CDH1, TNFRSF19, GM2A, FKBP5, PI3, PSPH, HORMAD1, SPRR2G ACTR2 and one or more selected from the group consisting of gene includes protein encoded from, said sensitive amount of protein in skin thereby a elevated relative to former characterized in that. Said protein expression dose compared to former from the skin susceptibility 2 to it is preferable that the.

Other embodiment: an sensitive skin or in the markers are diagnostic biochip, NFYA, MYOG ATF4 and one or more selected from the group consisting of transcription of the transcription factor gene includes protein encoded from, said sensitive amount of protein in skin thereby a elevated relative to former characterized in that. Said protein expression dose compared to former from the skin susceptibility 2 to it is preferable that the.

Another embodiment: an sensitive skin or in the markers are diagnostic biochip, KBTBD10, TTN, ACTA1, MYBPC1, MB, MYOZ1, TPM1, CA3, CASQ1, ENO3, RBP4, GYG2, PPP1 R1A, PYGM, G0S2, NEB, H19, LIPE, PLIN, H19, GPAM, FABP4, CIDEC, LPL, PCK1, ADIPOQ, PDE3B, ACVR1C, LPL, FABP4, DKFZP761N09121, TNMD, CRISP3 and ATP6V1B1, C8orf22 selected from the group consisting of from at least one gene includes protein encoded, said sensitive amount of protein is reduced relative to former from the skin is characterised in that it has a. Said protein expression dose susceptibility compared to former from the skin it is preferable that the reduced to 50% hereinafter.

Another embodiment: an sensitive skin or in the markers are diagnostic biochip, NFKB1, SRF, RELA, TCF3, POU2F1, USF JUN and one or more selected from the group consisting of an abrupt turning transcription of the transcription factor gene includes protein encoded from, said sensitive amount of protein is reduced relative to former from the skin is characterised in that it has a. Said protein expression dose susceptibility compared to former from the skin it is preferable that the reduced to 50% hereinafter.

1 or 2 of the present invention refers to in addition said table using polynucleotide of gene for diagnosis sensitive skin or in the provides diagnostic kit sensitive skin or in the.

One embodiment: an said sensitive skin diagnosis kit, HLA-C, TCEA1, IGHA1, TFRC, S100A8, LOC652128, SERPINB13, EHF, FLJ10781, TP63, CDH1, TNFRSF19, GM2A, FKBP5, PI3, PSPH, HORMAD1, SPRR2G ACTR2 and one or more selected from the group consisting of polynucleotides or a fragment thereof as gene, said fragment polynucleotide or more continuous 10 including one or more polynucleotides, or polynucleotides and said complementary polynucleotide fragments thereof; and a so as to obtain bonding of said polynucleotide includes a solid support, a hybridized polynucleotide said amount of transfer body and the former elevated relative to the sensitive skin or in the is characterized in that diagnosis. Said hybridized polynucleotides compared to and the former amount of transfer body 2 preferably is increased over back.

Other embodiment: an said sensitive skin diagnosis kit, NFYA, MYOG ATF4 and one or more selected from the group consisting of polynucleotides or a fragment thereof as gene, said fragment polynucleotide or more continuous 10 including one or more polynucleotides, or polynucleotides and said complementary polynucleotide fragments thereof; and a so as to obtain bonding of said polynucleotide includes a solid support, a hybridized polynucleotide said amount of transfer body and the former elevated relative to the sensitive skin or in the is characterized in that diagnosis. Said hybridized polynucleotides compared to and the former amount of transfer body 2 preferably is increased over back.

Another one embodiment: an said sensitive skin diagnosis kit, KBTBD10, TTN, ACTA1, MYBPC1, MB, MYOZ1, TPM1, CA3, CASQ1, ENO3, RBP4, GYG2, PPP1 R1A, PYGM, G0S2, NEB, H19, LIPE, PLIN, H19, GPAM, FABP4, CIDEC, LPL, PCK1, ADIPOQ, PDE3B, ACVR1C, LPL, FABP4, DKFZP761N09121, TNMD, CRISP3 and ATP6V1B1, C8orf22 selected from the group consisting of at least one gene as polynucleotides or a fragment thereof, said fragment polynucleotide or more continuous 10 including one or more polynucleotides, or polynucleotides and said complementary polynucleotide fragments thereof; and a so as to obtain bonding of said polynucleotide includes a solid support, a hybridized polynucleotide said amount of transfer body is reduced relative to and the former is sensitive skin or in the surface characterized in that diagnosis. Said polynucleotide a hybridized transcripts compared to amount and the former it is preferred that a this produces a 50% hereinafter.

Another one embodiment: an said sensitive skin diagnosis kit, NFYA, ATF4 and MYOG selected from the group consisting of at least one gene as polynucleotides or a fragment thereof, said fragment polynucleotide or more continuous 10 including one or more polynucleotides, or polynucleotides and said complementary polynucleotide fragments thereof; and a so as to obtain bonding of said polynucleotide includes a solid support, a hybridized polynucleotide said amount of transfer body is reduced relative to and the former is sensitive skin or in the surface characterized in that diagnosis. Said polynucleotide a hybridized transcripts compared to amount and the former it is preferred that a this produces a 50% hereinafter.

In said sensitive skin or in the diagnostic kit, probe as a Foley polynucleotides compared to skin insensitive increases or decreases is expressed in sensitive skin or in the gene for a marker includes (full length) or fragment thereof. The length of the fragments including 10 or more continuous it is preferred that a polynucleotide. 10 bps length of a probe which is non-specifically if less than is combined.

The present invention according to one embodiment: an said sensitive skin diagnosis kit, HLA-C, TCEA1, IGHA1, TFRC, S100A8, LOC652128, SERPINB13, EHF, FLJ10781, TP63, CDH1, TNFRSF19, GM2A, FKBP5, PI3, PSPH, HORMAD1, ACTR2 and SPRR2G selected from the group consisting of at least one gene polynucleotides fragments as polynucleotide constraint of 18-22 including one or more polynucleotides; and said gene polynucleotides of complementary polynucleotide fragments as constraint of 18-22 polynucleotide comprising a polynucleotide of which including one or more, said polynucleotides on the primers amplified DNA amount and the former elevated relative to the sensitive skin or in the is characterized in that diagnosis. Said polynucleotides amplified on the primers compared to DNA amount and the former 2 to it is preferable that the.

The present invention according to one embodiment: an said sensitive skin diagnosis kit, NFYA, ATF4 and MYOG selected from the group consisting of at least one gene polynucleotides fragments as polynucleotide constraint of 18-22 including one or more polynucleotides; and said gene polynucleotides of complementary polynucleotide fragments as constraint of 18-22 polynucleotide comprising a polynucleotide of which including one or more, said polynucleotides on the primers amplified DNA amount and the former elevated relative to the sensitive skin or in the is characterized in that diagnosis. Said polynucleotides amplified on the primers compared to DNA amount and the former 2 to it is preferable that the.

Another embodiment: an said sensitive skin diagnosis kit, KBTBD10, TTN, ACTA1, MYBPC1, MB, MYOZ1, TPM1, CA3, CASQ1, ENO3, RBP4, GYG2, PPP1 R1A, PYGM, G0S2, NEB, H19, LIPE, PLIN, H19, GPAM, FABP4, CIDEC, LPL, PCK1, ADIPOQ, PDE3B, ACVR1C, LPL, FABP4, DKFZP761N09121, TNMD, CRISP3 and ATP6V1B1, C8orf22 selected from the group consisting of at least one gene polynucleotides fragments as polynucleotide constraint of 18-22 including one or more polynucleotides; and said gene polynucleotides of complementary polynucleotide fragments as constraint of 18-22 polynucleotide comprising a polynucleotide of which including one or more, said polynucleotides amplified on the primers and the former amount DNA is sensitive skin or in the first and the second circular portions characterized in that diagnosis. Said polynucleotides amplified on the primers compared to DNA amount and the former it is preferable that the reduced to 50% hereinafter.

Another embodiment: an said sensitive skin diagnosis kit, NFKB1, SRF, RELA, TCF3, POU2F1, JUN and USF selected from the group consisting of at least one gene polynucleotides fragments as polynucleotide constraint of 18-22 including one or more polynucleotides; and said gene polynucleotides of complementary polynucleotide fragments as constraint of 18-22 polynucleotide comprising a polynucleotide of which including one or more, said polynucleotides amplified on the primers compared to DNA according to the decrease of the amount and the former is sensitive skin or in the characterized in that diagnosis. Said polynucleotides amplified on the primers compared to DNA amount and the former it is preferable that the reduced to 50% hereinafter.

Said polynucleotide encoded by the same or a sensitive skin or in the in diagnostic kit, polynucleotides as a Foley primer whose length is it is preferred that a personal 18-22. The, primer 18 bps length of and non-specifically bound if less than, greater than a self-associating each other primer surface 22 bps an electrode cost and productivity degradation in efficiency side do.

Furthermore, the present invention refers to said table 1 or 2 gene of polypeptides encoded by including monoclonal antibodies to provides diagnostic kit sensitive skin or in the.

In one in the embodiment, said sensitive skin diagnosis kit, HLA-C, TCEA1, IGHA1, TFRC, S100A8, LOC652128, SERPINB13, EHF, FLJ10781, TP63, CDH1, TNFRSF19, GM2A, FKBP5, PI3, PSPH, HORMAD1, ACTR2 and SPRR2G selected from the group consisting of at least one gene to polypeptides to be encoded by includes monoclonal antibodies against, said amount of antigen coupled to antibodies and the former elevated relative to the sensitive skin or in the is characterized in that diagnosis. Said amount of antigen coupled to antibodies compared to and the former 2 to it is preferable that the.

In other in the embodiment, said sensitive skin diagnosis kit, NFYA, ATF4 and MYOG selected from the group consisting of at least one gene to polypeptides to be encoded by includes monoclonal antibodies against, said amount of antigen coupled to antibodies and the former elevated relative to the sensitive skin or in the is characterized in that diagnosis. Said amount of antigen coupled to antibodies compared to and the former 2 to it is preferable that the.

In another in the embodiment, said sensitive skin diagnosis kit, KBTBD10, TTN, ACTA1, MYBPC1, MB, MYOZ1, TPM1, CA3, CASQ1, ENO3, RBP4, GYG2, PPP1 R1A, PYGM, G0S2, NEB, H19, LIPE, PLIN, H19, GPAM, FABP4, CIDEC, LPL, PCK1, ADIPOQ, PDE3B, ACVR1C, LPL, FABP4, DKFZP761N09121, TNMD, CRISP3 and ATP6V1B1, C8orf22 selected from the group consisting of at least one gene to polypeptides to be encoded by includes monoclonal antibodies against, said amount of antigen coupled to antibodies is reduced relative to and the former is sensitive skin or in the surface characterized in that diagnosis. Said amount of antigen coupled to antibodies compared to and the former it is preferable that the reduced to 50% hereinafter.

In another in the embodiment, said sensitive skin diagnosis kit, NFKB1, SRF, RELA, TCF3, POU2F1, JUN and USF selected from the group consisting of at least one gene to polypeptides to be encoded by includes monoclonal antibodies against, said amount of antigen coupled to antibodies is reduced relative to and the former is sensitive skin or in the surface characterized in that diagnosis. Said amount of antigen coupled to antibodies compared to and the former it is preferable that the reduced to 50% hereinafter.

The present invention refers to in addition said table 1 or encoded by gene 2 of protein and the uses and stimulate the winding provides method of screening for safeners.

In one in the embodiment, the method of screening safeners irritation said, HLA-C, TCEA1, IGHA1, TFRC, S100A8, LOC652128, SERPINB13, EHF, FLJ10781, TP63, CDH1, TNFRSF19, GM2A, FKBP5, PI3, PSPH, HORMAD1, ACTR2 and SPRR2G selected from the group consisting of at least one gene trial the proteins encoded by the substances; and said test substance of the protein is devices and confirming whether inhibitors action includes. Said protein action is inhibited and do not bind material test levels compared to a protein it is preferable that the reduced to 50% hereinafter.

In another in the embodiment, the method of screening safeners irritation said, KBTBD10, TTN, ACTA1, MYBPC1, MB, MYOZ1, TPM1, CA3, CASQ1, ENO3, RBP4, GYG2, PPP1 R1A, PYGM, G0S2, NEB, H19, LIPE, PLIN, H19, GPAM, FABP4, CIDEC, LPL, PCK1, ADIPOQ, PDE3B, ACVR1C, LPL, FABP4, DKFZP761N09121, TNMD, CRISP3 and ATP6V1B1, C8orf22 selected from the group consisting of at least one gene trial the proteins encoded by the substances; and enhance the action of this protein test substance said devices and confirming whether includes. Said protein action is inhibited and do not bind material test levels compared to a protein 2 times it is preferable that the enhanced.

Furthermore, the present invention refers to said gene gene transcription factor excluded from irritation using provides method of screening for safeners.

In one in the embodiment, the method of screening safeners irritation said, NFYA, MYOG ATF4 and at least one selected from the group consisting of transcription of the transcription factor gene promoter and including number 1 plasmid, and said transcription factor is a gene reporter sequences and DNA transfection step of agitating the mixed hot metal including number 2 plasmid into a cell, said step of treating the test substance cells, and said test substance the reporter devices and confirming whether inhibiting the expression of gene includes. Said test substance the reporter levels inhibited expression of gene test material compared to a protein and do not bind it is preferable that the reduced to 50% hereinafter.

In another in the embodiment, the method of screening safeners irritation said, NFKB1, SRF, RELA, TCF3, POU2F1, USF JUN and at least one selected from the group consisting of transcription of the transcription factor gene promoter and including number 1 plasmid, and said transcription factor is a gene reporter sequences and DNA transfection step of agitating the mixed hot metal including number 2 plasmid into a cell, said step of treating the test substance cells, and said test substance the reporter devices and confirming whether increasing the expression of gene includes. Said test substance the reporter levels inhibited expression of gene test material 2 compared to a protein and do not bind to it is preferable that the.

The 4 and 3 table a, expressed in sensitive skin or in the promoter gene, which increase transcription factor observed and the excessive (table 3) and a is reduced expression of transcription factor observed excessive promoter-gene (table 4) showed are.

said transcription factor gene, expressed in sensitive skin or in the gene, which increase promoter (table 1) is a transcription factor, observed and the excessive NFYA, MYOG and ATF4 and the like, with reduction of expression gene promoter (table 2) is a transcription factor, observed and the excessive NFKB1, SRF, RELA, TCF3, POU2F1, such as JUN and USF. Furthermore, gene expression with a selective ratio relative to the excessive simultaneously in a and reduced gene CEBPB include transcription factor observed. users can. Said transcription factor gene transcription regulation to a normal state the method screening for inhibitors irritation, an of the existing method of molecular biology techniques, thereby improving the efficiency of reporter gene, permits embodiment.

Said transcription factor gene using safeners winding and stimulate the method of screening for surfaces of a specifically to, were as follows..

General a universal promoter cultivation conditions and is also expressed in said table 3 or 4 combined with gene of plasmid number 1, and the transcription factor are combined which can activate the transfer DNA (Transcription Response Element) a reporter promoter having base:00 it ladles,glow of a firefly cathodically depositable coating compositions (firefly) combined with (luciferase) gene, use can be made of, plasmid number 2.

TRE sequences said number 2 plasmid, Matlnspector (Quandt et al. , 1995) and Transfac (Knuppel et al. , 1994) such as, for predicting (motif) motif, found program and analyzing gene accession ID of 1 and 2 of said table for inputting prioritized may then search for. COS7 cells with said transcription factor gene reporter-promoter TRE and the corresponding plasmid number 1 number 2, followed by transfection simultaneously plasmid, skin irritation time after the 24 drug candidate number inhibitors winding and for processing and cells base:00 it ladles, the cutting device is especially of measuring cathodically depositable coating compositions. Through, said candidate drug may be achieved, wherein an influence to activation of transcription factor said..

Expressed in sensitive skin or in the promoter gene, which increase transcription factor observed in base:00 it ladles, of cells as the active ingredient, at least in the case of cathodically depositable coating compositions when inhibitors sensitive skin effective takes short number. Vice versa, with reduction of expression of promoter-gene in the case of transcription factor observed excessive base:00 it ladles, of cells when cathodically depositable coating compositions for glyphosate having increased activity, inhibitors sensitive skin effective number the composition can be taken 2-2000. Furthermore, such, decreasing gene promoter in both in the case of transcription factor observed excessive auxiliary. may be utilized as screening method.

Also 1 and 2 through the. off at the first and the second aforementioned further the present invention.

NCBI Accession No reference altered was collecting promoter sequence of the gene. Ensembl genome database Upstream gene sequence from large, transfer disclosure site including genome region 2000 bp upstream from (transcription start site) was ensure sequence. MatInspector (Quandt et al. , 1995) and a Transfac (Knuppel et al. , 1994) (motif) motif such as for predicting, found, is opened for thereby implementing an analyzing transfer factor binding sites was retrieves (transcription factor binding site, TFBS). Search on the result, hyper geometry statistics method in, over-erased or appearance excessive transcription factor a Victory was selecting binding portions. Furthermore, selection that exceeds a preset expression changes gene cluster and valve cluster gene is expected to transcription factor between by extracting the by the coupling ring network control, was used to visible and graph. Figure 1 shows a also sensitive skin or in the skin tissue derived from expression compared to former insensitive in with a selective ratio relative to the gene promoters to and of the results of an-analysis, surface 2 derived from the sensitive skin or in the skin tissue in expression compared to former insensitive gene promoters is reduced to. of the results of an-analysis. Transcription factor is centered on that network and around the transcription factor adjustment of it is a target gene.

Hereinafter, the present invention through a in the embodiment described above but further, in the embodiment, or the like, that has to is to produce a rounding off on examples of the present invention, these category of the present invention only limited not.

Confirming gene marker sensitive skin or in the [in the embodiment 1]

1-1. Sampling RNA from tissue sample and of the tissue sample

Sensitive symptoms 9 9 a patient skin from name life and the control group patient took tissue sample. By biopsy are each tissue sample that were tissues at set. For purifying RNA from the tissue in, tree sol (trizol, Invitrogen, Carlsbad, CA, USA) after addition of fragmenting of cells, chloroform extended storage and is easily carried separated the RNA of supernatant. RNA of an upper layer obtained to enrich, isoforms of equal after the separation of the centrifugal into pro group glow , a precipitated RNA obtained. Precipitated RNA in order to remove the coating in the base was then being washed with ethanol 70%.

1-2. Experiment by

Total 54,675 species of human gene chip with integrated probe (transcripts) transfer body HG U133 Plus 2.0 (Affymetrix^®) to microarray experiment is performed for all the. According to sensitivity skin change in of expression gene expressing therefor expression pattern has been confirmed. 10 ug of RNA using a cDNA synthesis, in vitro have been synthesised (Biotinylated) cRNA biotinylated in a transfer method.

Specifically, Affymetrix along the protocol by a special arrangement, first the T7 primers and including cDNA using reverse transcriptase have been synthesised RNA from a promoter. Furthermore, double-stranded cDNA a circular by a use of the reaction takes place in the transfer in vitro have been synthesised a cRNA. In vitro transfer reaction is, (labeling) for amplification and cRNA for labeling ribonucleic biotinylated polymerase and T7 RNA by using nucleotide analogs.

The thus produced planar metallised textile structure biotinylated cRNAs has, or a fragment thereof of (fragmentation) after, said gene chip 42 °C time been hybridization in 17. Said gene chip a cleaning, obviated and post dyeing scanning the n bit parallel data inputted, data pretreatment Global scaling a algorithm is (normalization) normalization basic software GenPlex adapted for use in high-density.

As a result, the control group samples and sensitive skin or in the gene expression in sample with a gene group show differences significantly p-value and a gene expression amount was accomplished compared width.

Based on the gene expression pattern, in the case of reduced expression gene gene at a 50% hereinafter, is upregulated a gene in the case of increased times 2 selects sensitive skin or in the gene used for diagnosing may be utilized a gene extracted this marker as a basis. The result thereof is said table 1 and 2. are is additionally prepared a gene.