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NOVEL PROTEIN IDENTIFICATION METHOD USING YEAST AND FRAGMENTED CDNA LIBRARY NEW

外国特許コード F170009304
整理番号 (S2016-0677-N0)
掲載日 2017年12月20日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2017JP020614
国際公開番号 WO 2017209280
国際出願日 平成29年6月2日(2017.6.2)
国際公開日 平成29年12月7日(2017.12.7)
優先権データ
  • 特願2016-112107 (2016.6.3) JP
発明の名称 (英語) NOVEL PROTEIN IDENTIFICATION METHOD USING YEAST AND FRAGMENTED CDNA LIBRARY NEW
発明の概要(英語) The purpose of the present invention is to provide a novel method for identifying a protein in a yeast nucleus. More specifically, the present invention relates to a protein identification method which comprises: in a yeast nucleus, expressing a fusion protein comprising a nuclear localization signal, a DNA-binding protein and a bait protein, and another fusion protein comprising a nuclear localization signal, a transcriptional activation protein and a prey protein encoded by a fragmented partial cDNA, said prey protein lacking a signal peptide, a cell membrane or intracellular organelle localization sequence or a transmembrane domain; and then detecting the binding of the bait protein to the prey protein using the activity, luminescence, etc. of a reporter gene as an indication.
特許請求の範囲(英語) [claim1]
1. The process which cultures the yeast transformant which introduces the 1st and 2nd vector is included, being identification method of the protein,
As for the 1st vector, the gene which the nuclear movement signal the cord/code is done and the DNA connection protein the cord/code is done the gene and the gene which the decoy protein the cord/code is done implication,
The 2nd vector to include with the gene which the nuclear movement signal the cord/code is done and the copying activated protein the cord/code is done the gene and the part cDNA which is fragmented, the said as for the part cDNA where it fragments, signal peptide, the cell membrane or small organ localization arrangement or the cell membrane penetration territory inside the cell to be something which the game protein which deficiency is done the cord/code is done, at the same time,
The fusion protein which includes with the game protein which the cord/code is done reveals the aforementioned yeast transformant in, the aforementioned nuclear movement signal and the fusion protein, and the aforementioned nuclear movement signal and the copying activated protein which include the DNA connection protein and the decoy protein with the part cDNA which is fragmented nuclear, connection with the said decoy protein and the said game protein, detects the activity of the reporter gene in index,
The aforementioned method.
[claim2]
2. The decoy protein is the ligand, at the same time the game protein is the receptor protein, method of claim 1 statement.
[claim3]
3. It is the protein which the cord/code is done with the part cDNA where the decoy protein excludes various localized signal arrangements, method of claim 1 statement.
[claim4]
4. CDNA which could by random prime reversal copying, possesses length 100-1800bp.
[claim5]
5. Length is 100-1000bp, cDNA of claim 4 statement.
[claim6]
6. The gene and the cord/code it does the copying activated protein the gene and part cDNA which is fragmented being the vector which includes, the said the part cDNA which is fragmented claim with 4 where the cord/code it does the nuclear movement signal or is cDNA of 5 statements, the aforementioned vector.
[claim7]
7. Claim cDNA of 4 or 5 statements or the vector of claim 6 statement is included, either of the claim 1-3 the reagent kit for identification method of the protein of 1 section statements.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • KAGOSHIMA UNIVERSITY
  • 発明者(英語)
  • KISHIDA SHOSEI
  • KOYAMA HIROFUMI
  • KISHIDA MICHIKO
  • IIJIMA MIKIO
国際特許分類(IPC)
指定国 (WO2017209280)
National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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