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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.   Supplementary Information  The online version contains supplementary material available at 10.1186/s13036-021-00270-9.

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