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METHOD FOR IDENTIFYING BIOACTIVE PROTEIN, AND BIOACTIVE PROTEIN OBTAINED BY SAID METHOD

外国特許コード F170009076
整理番号 (S2015-1988-N64)
掲載日 2017年5月29日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2016JP077198
国際公開番号 WO 2017047672
国際出願日 平成28年9月14日(2016.9.14)
国際公開日 平成29年3月23日(2017.3.23)
優先権データ
  • 特願2015-180831 (2015.9.14) JP
発明の名称 (英語) METHOD FOR IDENTIFYING BIOACTIVE PROTEIN, AND BIOACTIVE PROTEIN OBTAINED BY SAID METHOD
発明の概要(英語) The purpose of the present invention is to provide a method for identifying a bioactive protein translated from an open reading frame (ORF) other than a mORF present in mRNA. This problem is solved by a method for identifying a bioactive protein that is characterized by identifying a bioactive protein-encoding ORF other than the main ORF in the mRNA of a eukaryote, said method including: (1) a step in which an expression vector containing a candidate ORF is introduced into a cell, and the vector-introduced cell is cultured; (2) a step in which a protein that binds to a candidate protein, which is translated from the candidate ORF, is detected by immunoprecipitating the candidate protein from the cultured cell; and (3) a step in which the candidate protein, for which another protein that binds to the candidate protein was detected, is determined to be a bioactive protein.
特許請求の範囲(英語) [claim1]
1. The open leading frame of main in mRNA of the eucaryote (ORF) ORF which the protein which possesses the physiological activity other than the cord/code has been done being identification method of the physiological active protein which features that it specifies,
Being the process which cultures the cell which (1) introduces the manifestation vector which installs candidacy ORF into the cell, is introduced, the start codon of the aforementioned candidacy ORF, is non AUG where one base of AUG or AUG is the other base,
(2) from the cultured cell, the process which detects the protein which is connected to the candidacy protein which is translated from candidacy ORF,
(3) the process which as the physiological active protein decides the candidacy protein where the other protein which is connected to the candidacy protein is detected,
Identification method of the physiological active protein which is included.
[claim2]
2. In the aforementioned cell culture process (1), the [puroteasomu] inhibiter or the lysosome inhibiter is added in the cell culture solution, in claim 1 identification method of the physiological active protein of statement.
[claim3]
3. The aforementioned connection protein detection process (2) before, revelation of the candidacy protein which is translated beforehand from candidacy ORF is detected, the process which selects the candidacy ORF where the revelation quantity is many is included, in claim 1 or 2 identification method of the physiological active protein of statement.
[claim4]
4. The aforementioned candidacy ORF,
(A) The start codon of candidacy ORF, the start codon of the aforementioned main ORF compared to 5 ' exists on UTR side,
(B) The cord/code of candidacy ORF the protein which is done is 10 amino acids or more,
(C) Candidacy ORF, it is ORF other than main ORF in mRNA of the eucaryote of 1 kinds, the amino acid arrangement which is translated from the aforementioned candidacy ORF, it is ORF which possesses the identity of 50% or more vis-a-vis the amino acid arrangement which is translated from ORF other than main ORF in mRNA of the eucaryote above other 1 kinds or 2 kinds,
(D) The start codon of candidacy ORF being AUG, and
(E) It is ORF where the amino acid arrangement which agrees from the database of candidacy ORF making use of the molecular weight of the peptide which analyzes the peptide which has been revealed in the cell of the eucaryote with mass spectrometry, is obtained is searched,
The condition of one or more which is selected from the group which consists of is satisfied, either of the claim 1-3 in one section identification method of the physiological active protein of statement.
[claim5]
5. In the aforementioned connection protein detection process (2), the method of detecting the aforementioned connection protein, is the method of being selected from the group which consists of immunity sinkage, yeast two hybrid method, protein array method, label method, and the BioID method which use the peroxidase, either of the claim 1-4 in one section identification method of the physiological active protein of statement.
[claim6]
6. The aforementioned eucaryote, is Mammalia, either of the claim 1-5 in one section identification method of the physiological active protein of statement.
[claim7]
7. (1) the protein which consists of the amino acid arrangement which is displayed with arrangement number 1,
The protein which shows the function which controls the abnormality of the spiritual activity which (2) includes the amino acid arrangement which is displayed with arrangement number 1, furthermore, becomes the basis of childcare conduct,
The protein which shows the function which controls the abnormality of the spiritual activity where 1 or several amino acids are deficient (3) at the time of amino acid arranging which is displayed with arrangement number 1, substitution and insertion, and/or consist of the amino acid arrangement which is added, furthermore, become the basis of childcare conduct, or,
The protein which shows the function which controls the abnormality of the spiritual activity where 1 or several amino acids are deficient (4) at the time of amino acid arranging which is displayed with arrangement number 1, substitution and insertion, and/or include the amino acid arrangement which is added, furthermore, become the basis of childcare conduct.
[claim8]
8. (1) the protein which consists of the amino acid arrangement which is displayed with arrangement number 10 or 11,
The protein which (2) is displayed with arrangement number 10 or 11, includes the amino acid arrangement which furthermore, possesses the connection talent of Peroxiredoxin 1,
The protein where 1 or several amino acids are deficient (3) at the time of amino acid arranging which is displayed with arrangement number 10 or 11, substitution and insertion, and/or is added, consist of the amino acid arrangement which furthermore, possess the connection talent of Peroxiredoxin 1, or,
The protein where 1 or several amino acids are deficient (4) at the time of amino acid arranging which is displayed with arrangement number 10 or 11, substitution and insertion, and/or is added, include the amino acid arrangement which furthermore, possess the connection talent of Peroxiredoxin 1.
[claim9]
9. (1) the protein which consists of the amino acid arrangement which is displayed with arrangement number 14 or 15,
The protein which (2) is displayed with arrangement number 14 or 15, includes the amino acid arrangement which furthermore, possesses the connection talent of Q Subcomponent Binding Protein,
The protein where 1 or several amino acids are deficient (3) at the time of amino acid arranging which is displayed with arrangement number 14 or 15, substitution and insertion, and/or is added, consist of the amino acid arrangement which furthermore, possess the connection talent of Q Subcomponent Binding Protein, or,
The protein where 1 or several amino acids are deficient (4) at the time of amino acid arranging which is displayed with arrangement number 14 or 15, substitution and insertion, and/or is added, include the amino acid arrangement which furthermore, possess the connection talent of Q Subcomponent Binding Protein.
[claim10]
10. Either of the claim 7-9 in one section the polynucleotide which the protein of statement the cord/code is done.
[claim11]
11. The manifestation vector which includes the polynucleotide of statement in claim 10.
[claim12]
12. The transformant which includes the polynucleotide of statement in claim 10.
[claim13]
13. Either of the claim 7-9 the antibody which is connected to the protein of statement in one section or the fragment.
[claim14]
14. Revelation of the gene which the protein of statement the cord/code is done part in claim 7 or the knockout non human animal or the cell which is controlled completely.
[claim15]
15. Screening method of the chemical compound which controls the abnormality of the spiritual activity which features that candidacy chemical compound is prescribed to the knockout non human animal or the cell of statement, in claim 14, becomes the basis of childcare conduct.
  • 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
  • TOKYO INSTITUTE OF TECHNOLOGY
  • LIFEIS INC.
  • 発明者(英語)
  • AIZAWA YASUNORI
  • KITANO SHOHEI
  • KIDA YUICHIRO
国際特許分類(IPC)
指定国 (WO201747672)
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 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 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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