公开(公告)号：US20110183402A1

公开(公告)日：2011-07-28

申请号：US13070583

申请日：2011-03-24

申请人： JAMES M. CREGG ,  JOHN LATHAM ,  MARK LITTON ,  RANDALL SCHATZMAN ,  IIYA I. TOLSTORUKOV

发明人： JAMES M. CREGG ,  JOHN LATHAM ,  MARK LITTON ,  RANDALL SCHATZMAN ,  IIYA I. TOLSTORUKOV

IPC分类号： C12N1/19

CPC分类号： C07K16/2896 ,  C07K16/00 ,  C07K16/18 ,  C07K16/2809 ,  C07K2317/24 ,  C12N15/81 ,  C12N15/815

摘要： Methods are provided for the synthesis and secretion of recombinant proteins preferably large mammalian proteins or hetero-multimeric proteins at high levels and for prolonged time in polyploid, preferably diploid yeast. These methods use various mating competent yeast, including Pichia. In a preferred embodiment, a first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

摘要翻译： 提供了用于在多倍体，优选二倍体酵母中以高水平合成和分泌重组蛋白，优选大量哺乳动物蛋白或杂多聚体蛋白并延长时间的方法。 这些方法使用各种交配的感受态酵母，包括毕赤酵母。 在优选的实施方案中，将第一表达载体转化为第一单倍体细胞; 并将第二表达载体转化成第二单倍体细胞。 鉴定转化的单倍体细胞，每个细胞各自合成不相同的多肽，然后遗传交叉或融合。 所得的二倍体菌株用于产生和分泌完全组装和生物功能的杂多聚体蛋白。