Rapid monitoring of the target protein expression with a fluorescent signal based on a dicistronic construct in Escherichia coli.

ABSTRACT: Real-time quantification of recombinant proteins is important in studies on fermentation engineering, cell engineering, etc. Measurement of the expression level of heterologous proteins in bacterial fermentation broth has traditionally relied on time-consuming and labor-intensive procedures, such as polyacrylamide gel electrophoresis, immunoblot analysis, and biological activity assays. We describe a simple, fast, and high sensitive assay for detecting heterologous proteins production in bacteria either at the overall level (fluorescence spectrophotometry) or at the individual level (fluorescence microscopic image) in this study. Based on a dicistronic model, the translation of target gene in the upstream open reading frame (ORF) was coupled with the synthesis of the mCherry reporter in th

SUBMITTER: Hui CY 

PROVIDER: S-EPMC5962521 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature