发明名称 |
DNA, cell cultures and methods for the secretion of heterologous proteins and periplasmic protein recovery. |
摘要 |
Direct linkage of DNA encoding prokaryotic signals such as E. coli enterotoxin with DNA encoding mature eukaryotic proteins, followed by transformation and culture of bacterial hosts characterized by a periplasmic space, yields substantial amounts of periplasmic mature protein. Constitutively promoted gram negative bacterial mutants are transformed with vectors containing DNA encoding a secretable heterologous protein under the transcriptional control of a constitutively promoted promoter. Periplasmic proteins are recovered from transformed gram negative bacteria by a process comprising freezing and thawing the cells. Advantages are obtained by culturing the cells in phosphate-limiting media and by killing the cells prior to separation of periplasmic proteins.
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申请公布号 |
EP0177343(A1) |
申请公布日期 |
1986.04.09 |
申请号 |
EP19850307044 |
申请日期 |
1985.10.02 |
申请人 |
GENENTECH, INC. |
发明人 |
BOCHNER, BARRY RONALD;CHANG, CHUNG-NAN;GRAY, GREGORY LAWRENCE;HEYNEKER, HERBERT LOUIS;MCFARLAND, NANCY CHADWICK;OLSON, KENNETH CHARLES;PAI, RONG-CHANG;REY, MICHAEL WILLARD |
分类号 |
C12N1/21;C07K14/24;C07K14/61;C12N1/00;C12N5/00;C12N15/00;C12N15/09;C12N15/62;C12N15/70;C12P21/02;C12R1/19;(IPC1-7):C12N15/00;C07K13/00;C12P21/00;C07K15/00;C07H21/04 |
主分类号 |
C12N1/21 |
代理机构 |
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代理人 |
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地址 |
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