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METHODS FOR PRODUCING HETEROLOGOUS POLYPEPTIDES IN THIOL-DISULFIDE OXIDOREDUCTASE-DEFICIENT BACTERIAL MUTANT CELLS
METHODS FOR PRODUCING HETEROLOGOUS POLYPEPTIDES IN THIOL-DISULFIDE OXIDOREDUCTASE-DEFICIENT BACTERIAL MUTANT CELLS
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机译:硫醇二硫化物氧化还原酶缺陷型细菌突变细胞中异源多肽的生产方法
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摘要
The present invention relates to methods of producing a heterologous polypeptide, comprising: (a) cultivating a mutant of a parent bacterial cell in a medium for the production of the heterologous polypeptide, wherein (i) the mutant cell comprises a first polynucleotide encoding the heterologous polypeptide which comprises two or more (several) cysteines, and a second polynucleotide comprising a modification of a gene encoding a thiol-disulfide oxidoreductase that incorrectly catalyzes the formation of one or more (several) disulfide bonds between the two or more (several) cysteines of the heterologous polypeptide, and (ii) the mutant cell is deficient in production of the thiol-disulfide oxidoreductase compared to the parent bacterial cell when cultivated under the same conditions; and (b) recovering the heterologous polypeptide from the cultivation medium. The present invention also relates to such bacterial mutants and methods for producing such bacterial mutants.
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