Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance.
Microb Cell Fact
; 20(1): 13, 2021 Jan 11.
Article
em En
| MEDLINE
| ID: mdl-33430875
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 downregulation. The over-expression process is bactericidal and is observed for both soluble and membrane proteins.CONCLUSIONS:
We demonstrate that it is the act of high-level over-expression of exogenous proteins in E. coli that sets in motion a chain of events leading to > 99.9% cell death. These results redefine our understanding of protein over-production and link it to the adaptive survival response seen in the development of antimicrobial resistance.Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Adaptação Fisiológica
/
Proteínas de Escherichia coli
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Escherichia coli
/
Proteínas de Membrana
Idioma:
En
Revista:
Microb Cell Fact
Assunto da revista:
BIOTECNOLOGIA
/
MICROBIOLOGIA
Ano de publicação:
2021
Tipo de documento:
Article
País de afiliação:
Reino Unido