Application of the E-coli trp promoter

被引:18
作者
Bass, SH
Yansura, DG
机构
[1] Genentech Inc, Dept Mol Biol, San Francisco, CA 94080 USA
[2] Maxygen Inc, Redwood City, CA 94063 USA
关键词
Escherichia coli; tryptophan promoter; gene expression; protein production;
D O I
10.1385/MB:16:3:253
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The Escherichia coli tryptophan (trp) promoter has been used extensively for the high level production of proteins on a small and large scale. This regulated promoter is readily available, relatively easy to turn on, and can be used in essentially any E. coli host background. This article gives a detailed use of the try promoter including the design of expression vectors, subsequent culture conditions for promoter induction, and, finally, a protocol for the most common way of detecting the newly synthesized protein of interest. Its successful use for heterologous protein expression, however, sometimes requires consideration of parameters other than transcription such as translation initiation, translation elongation, and proteolysis. In this respect we offer guidance in getting through these post-transcriptional problems, which can occur with the use of any promoter.
引用
收藏
页码:253 / 260
页数:8
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