Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance.

ABSTRACT:

Background

The E. coli pET system is the most widely used protein over-expression system worldwide. It relies on the assumption that all cells produce target protein and it is generally believed that integral membrane protein (IMP) over-expression is more toxic than their soluble counterparts.

Results

Using GFP-tagged proteins, high level over-expression of either soluble or IMP targets results in > 99.9% cell loss with survival rate of only < 0.03%. Selective pressure generates three phenotypes: large green, large white and small colony variants. As a result, in overnight cultures, ~ 50% of the overall cell mass produces no protein. Genome sequencing of the phenotypes revealed genomic mutations that causes either the loss of T7 RNAP activity or its transcriptional downregul

SUBMITTER: James J 

PROVIDER: S-EPMC7798265 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature