公开(公告)号：US06841361B1

公开(公告)日：2005-01-11

申请号：US09959548

申请日：2000-04-26

申请人： Shusaku Oka ,  Seiji Sato ,  Naohiko Higashikuni ,  Masaaki Kondo ,  Toshiyuki Kudo ,  Shigeaki Watanabe ,  Yoshihiro Waki ,  Hirotaka Yuki

发明人： Shusaku Oka ,  Seiji Sato ,  Naohiko Higashikuni ,  Masaaki Kondo ,  Toshiyuki Kudo ,  Shigeaki Watanabe ,  Yoshihiro Waki ,  Hirotaka Yuki

IPC分类号： C12N15/09 ,  C07K14/575 ,  C07K14/62 ,  C07K19/00 ,  C12N1/21 ,  C12N15/00 ,  C12N15/17 ,  C12P21/02 ,  C12R1/08 ,  C12P21/04

CPC分类号： C07K14/62 ,  C07K14/575 ,  C07K2319/00

摘要： This invention relates to a DNA encoding a fusion protein of formula (I) [Y]-[X1]-[B-chain]-[X2]-[Linker]-[X3]-[A-chain]  (I) wherein Y is a leader peptide sequence for the expression and secretion of the protein, comprising at least one amino acid residue; X1 is an amino acid sequence which is cleavable enzymatically or chemically; B-chain is the amino acid sequence of insulin B chain; X2 is an amino acid sequence which is cleavable enzymatically; Linker is a linker sequence comprising at least one amino acid residue; X3 is an amino acid sequence which is cleavable enzymatically; and A- chain is the amino acid sequence of insulin A chain, and to a method for producing insulin using an expression system containing the DNA in high efficiency and high yield.

摘要翻译： 本发明涉及编码式（I）的融合蛋白的DNA，其中Y是蛋白质的表达和分泌的前导肽序列，其包含至少一个氨基酸残基; X1是可酶切或化学切割的氨基酸序列 B链是胰岛素B链的氨基酸序列; X2是可酶切的氨基酸序列;接头是包含至少一个氨基酸残基的接头序列; X3是可酶切的氨基酸序列; A链是胰岛素A链的氨基酸序列，以及使用含有高效率和高收率的DNA的表达系统来生产胰岛素的方法。

公开(公告)号：US07037684B2

公开(公告)日：2006-05-02

申请号：US10221677

申请日：2001-03-13

申请人： Shigezo Udaka ,  Seiji Sato ,  Toshiyuki Kudo ,  Shusaku Oka ,  Naohiko Higashikuni ,  Masaaki Kondo

发明人： Shigezo Udaka ,  Seiji Sato ,  Toshiyuki Kudo ,  Shusaku Oka ,  Naohiko Higashikuni ,  Masaaki Kondo

IPC分类号： C07H21/04 ,  C12P21/04 ,  C12N1/20 ,  C12N15/00 ,  A61K38/28

CPC分类号： C12P21/02 ,  C07K14/32 ,  C07K14/62 ,  C07K2319/02 ,  C07K2319/035 ,  C07K2319/75 ,  C12N9/90 ,  C12N15/625 ,  C12N15/75

摘要： The invention relates to a DNA that enables co-expression of a protein disulfide isomerase and a polypeptide having disulfide bonds. The invention further relates to a process of producing polypeptides using the DNA and a process of enhancing the efficiency of formation of correct disulfide bond in an expression system of a gene recombinant polypeptide having disulfide bonds.

摘要翻译： 本发明涉及能够共表达蛋白质二硫键异构酶和具有二硫键的多肽的DNA。 本发明还涉及使用该DNA产生多肽的方法和在具有二硫键的基因重组多肽的表达系统中提高正确二硫键形成的效率的方法。

公开(公告)号：US06506595B2

公开(公告)日：2003-01-14

申请号：US09280030

申请日：1999-03-26

申请人： Seiji Sato ,  Naohiko Higashikuni ,  Toshiyuki Kudo ,  Masaaki Kondo

发明人： Seiji Sato ,  Naohiko Higashikuni ,  Toshiyuki Kudo ,  Masaaki Kondo

IPC分类号： C12N1500

CPC分类号： C12N15/625 ,  C07K14/605 ,  C07K14/61 ,  C07K14/62 ,  C07K14/655 ,  C07K2319/034 ,  C07K2319/21 ,  C07K2319/50 ,  C07K2319/75

摘要： This invention relates to a DNA comprising a nucleotide sequence encoding a fusion protein, wherein the fusion protein comprises: a sequence of signal peptide for Bacillus cell wall protein (CWP); a tag sequence for separation and purification of the fusion protein; a linker sequence; a sequence for chemical or enzymatic cleavage; and an exogenous polypeptide sequence, the sequences being linked in order, the signal peptide, tag and linker being optional sequences; and wherein the nucleotide sequence encoding a fusion protein is ligated to 3′-end of a nucleic acid sequence comprising a Bacillus promoter region; to a vector comprising the DNA; to a bacterium belonging to the genus Bacillus comprising the vector; and to a process for preparation of a useful polypeptide by culture of the bacterium.

摘要翻译： 本发明涉及包含编码融合蛋白的核苷酸序列的DNA，其中融合蛋白包含：用于芽孢杆菌细胞壁蛋白（CWP）的信号肽序列; 用于分离和纯化融合蛋白的标签序列; 接头序列; 用于化学或酶切割的序列; 和外源多肽序列，序列依次连接，信号肽，标签和接头是任选的序列; 并且其中将编码融合蛋白的核苷酸序列连接到包含芽孢杆菌启动子区的核酸序列的3'-末端; 涉及包含该DNA的载体; 涉及属于包含载体的芽孢杆菌属的细菌; 以及通过细菌培养制备有用多肽的方法。