发明授权
EP0931160B1 MUTATION DETECTION USING PEPTIDE TAGGED IN VITRO SYNTHESIZED PROTEINS
失效
突变借助体外检测中合成的蛋白质用肽MARK
- 专利标题: MUTATION DETECTION USING PEPTIDE TAGGED IN VITRO SYNTHESIZED PROTEINS
- 专利标题(中): 突变借助体外检测中合成的蛋白质用肽MARK
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申请号: EP96931558.9申请日: 1996-09-13
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公开(公告)号: EP0931160B1公开(公告)日: 2008-12-17
- 发明人: Garvin, Alex M.
- 申请人: Garvin, Alex M.
- 申请人地址: 13, rue du Chemin de Fer 68480 Durmenach FR
- 专利权人: Garvin, Alex M.
- 当前专利权人: Garvin, Alex M.
- 当前专利权人地址: 13, rue du Chemin de Fer 68480 Durmenach FR
- 代理机构: Kirkham, Nicholas Andrew
- 国际公布: WO1998011249 19980319
- 主分类号: C12P19/34
- IPC分类号: C12P19/34 ; G01N33/543 ; C12Q1/68
摘要:
A process for detecting protein altering mutations in genes. Coding sequence of the gene is PCR amplified with a 5' primer that contains at its 5' end a polymerase binding site, a translation initiation site, and an in frame sequence coding for a peptide tag, followed by the in frame 5' end of the test sequence. After PCR amplification of the test sequence, the PCR product is used as template to make mRNA in an in vitro transcription reaction using an RNA polymerase that recognizes the polymerase binding site incorporated into the 5' PCR primer. The mRNA is then used as template to make protein in an in vitro translation reaction. The protein encoded by the test sequence has at its amino terminus a peptide tag that can be used to either detect the protein, or to purify the protein for further analysis.
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