Facile promoter deletion in Escherichia coli in response to leaky expression of very robust and benign proteins from common expression vectors.

ABSTRACT:

Background

Overexpression of proteins in Escherichia coli is considered routine today, at least when the protein is soluble and not otherwise toxic for the host. We report here that the massive overproduction of even such "benign" proteins can cause surprisingly efficient promoter deletions in the expression plasmid, leading to the growth of only non-producers, when expression is not well repressed in the newly transformed bacterial cell. Because deletion is so facile, it might impact on high-throughput protein production, e.g. for structural genomics, where not every expression parameter will be monitored.

Results

We studied the high-level expression of several robust non-toxic proteins using a T5 promoter under lac operator control. Full induction leads to no significant g

SUBMITTER: Kawe M 

PROVIDER: S-EPMC2655282 | biostudies-literature | 2009 Jan

REPOSITORIES: biostudies-literature