发明公开
- 专利标题: PRODUCTION DE PLASMIDES ET EXPRESSION DE PROTEINES RECOMBINANTES DANS DES CELLULES CULTIVEES SANS ANTIBIOTIQUES
- 专利标题(英): Plasmid production and expression of recombinant proteins in cells cultivated without antibiotics
- 专利标题(中): 质粒生产和表达的重组蛋白在没有抗生素的培养细胞
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申请号: EP08787440.0申请日: 2008-08-22
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公开(公告)号: EP2195434A1公开(公告)日: 2010-06-16
- 发明人: SCHERMAN, Daniel , MARIE, Corinne
- 申请人: Centre National de la Recherche Scientifique (CNRS) , UNIVERSITE PARIS DESCARTES , Institut National De La Sante Et De La Recherche Medicale
- 申请人地址: 3, rue Michel-Ange 75016 Paris FR
- 专利权人: Centre National de la Recherche Scientifique (CNRS),UNIVERSITE PARIS DESCARTES,Institut National De La Sante Et De La Recherche Medicale
- 当前专利权人: Centre National de la Recherche Scientifique (CNRS),UNIVERSITE PARIS DESCARTES,Institut National De La Sante Et De La Recherche Medicale
- 当前专利权人地址: 3, rue Michel-Ange 75016 Paris FR
- 代理机构: Warcoin, Jacques
- 优先权: FR0757175 20070824
- 国际公布: WO2009027351 20090305
- 主分类号: C12N15/63
- IPC分类号: C12N15/63 ; C12N15/64 ; C12N15/65
摘要:
The invention relates to a mutated cell such as a bacteria or yeast in which the thyA gene coding thymidylate synthase includes a nonsense codon, preferably the amber codon, said nonsense codon replacing a codon coding an amino acid and inducing the interruption of thyA gene translation and the auxotropy of the cell for the thymidine. Advantageously, the endA gene coding the endonuclease 1 and/or the recA gene coding the recombinase is inactivated in said mutated cell. The invention also relates to an expression plasmid including a transgene and a sequence of a suppressing ARM structural gene containing an anticodon that can be paired with the nonsense codon of the thyA gene and is specific of an amino acid capable of restoring the translation of the mutated thyA gene and thereby obtaining a protein of the wild or mutated type having a thymidylate synthase activity. The invention also relates to a method for the multiplication of the expression plasmid.
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