원문정보
초록
영어
E. coli is one of the most preferred strains for the production of recombinant proteins because of many reasons, such as its fast growth rate and many available genetic tools. Still, there are several problems in using E. coli strain as a microbial protein factory. Formation of inclusion body is one of problems to overcome, which is frequently observed when the recombinant proteins are highly expressed. We hypothesized that the knockout of a specific gene could
reduce the formation of inclusion body through direct or indirect impacts on protein synthesis rate in E. coli. As such, we investigated the relationships between a specific genotype and inclusion body forming phenotypes. Specifically, we transformed a plasmid containing a reporter protein (GFP) under the control of a strong inducible promoter into a systematic knockout library. The
transformed knockout library were screened by FACS and microplate reader. Based on the intensity of the fluorescence of GFP, we selected knockout mutants which accumulated higher amount of soluble GFP as compared to the parent strain. Accumulation of higher amounts of soluble GFP in knockout mutant was confirmed using SDS-PAGE. These results suggest that the single knockout of the specific gene can affect the formation of inclusion body in E.
coli.