CRISPR/Cas9 mediated T7 RNA polymerase gene knock-in in E. coli BW25113 makes T7 expression system work efficiently

Abstract 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 which the RNA Polymerase gene resides chromosome. In study, we successfully introduced chromosomal copy under lacUV5 promoter into Escherichia coli BW25113. The worked efficiently mutant strain named BW25113-T7. We demonstrated that could satisfactorily produce 5-Aminolevulinic Acid via C5 pathway. A final study was designed to enhance controllability by constructing Promoter Variants Library. These efforts advanced E. BW25113-T7 be practical host for future metabolic engineering efforts.