A novel system for stable, high-level expression from the T7 promoter.

ABSTRACT:

Background

The most widespread, efficient prokaryotic protein-producing system is one where the T7 phage polymerase recognizes the T7 phage promoter (T7 p/p system). Unfortunately, in this system, target protein expression gradually declines and is often undetectable following 3 to 5 subcultures. Although a number of studies have attempted to stabilize the expression levels of the T7 p/p system, none has resolved the problem adequately and thus precludes the use of this system for the production of recombinant proteins on a large scale.

Results

We created an expression cassette enabling stable, high-level expression in the T7p/p system. The cassette was tested with two different vector backbones and two target proteins. In all experiments, the expression system using the new

SUBMITTER: Kesik-Brodacka M 

PROVIDER: S-EPMC3512507 | biostudies-literature | 2012 Aug

REPOSITORIES: biostudies-literature