CRISPR/Cas9 mediated T7 RNA polymerase gene knock-in in E. coli BW25113 makes T7 expression system work efficiently
发布于: 2023-08-28 来源: 国家饲料工程技术研究中心
关键词：E. coli, CRISPR/cas9, T7 Expression System, Fluorescent Protein, 5-Aminolevulinic Acid, promoter variants
摘要：T7 Expression System is a common method of ensuring tight control and high-level induced expression. However, this
system can only work in some bacterial strains in which the T7 RNA Polymerase gene resides in the chromosome. In
this study, we successfully introduced a chromosomal copy of the T7 RNA Polymerase gene under control of the lacUV5
promoter into Escherichia coli BW25113. The T7 Expression System worked efficiently in this mutant strain named
BW25113-T7. We demonstrated that this mutant strain could satisfactorily produce 5-Aminolevulinic Acid via C5
pathway. A final study was designed to enhance the controllability of T7 Expression System in this mutant strain by
constructing a T7 Promoter Variants Library. These efforts advanced E. coli BW25113-T7 to be a practical host for future
metabolic engineering efforts.