Design and Application of a Cumate-inducible Promoter for Corynebacterium glutamicum
[Objective]Corynebacterium glutamicum is an important industrial microorganism,and its metabolic engineering modification by gene editing may effectively broaden the diversity of its fermentation products.The lack of high-intensity,low-leakage,and low-cost inducible promoters limits the metabolic engineering modification,thereby the design and application of novel inducible promoters are necessary.[Method]An cumate-induced promoter PH10-CuO was constructed by embedding the operator sequence CuO on the constitutive promoter PH10.[Result]Using green fluorescent protein as a quantitative reporter,the relative fluorescence intensity was low due to the basal leakage of PH10-CuO when without the inducer 4-isopropylbenzoic acid;and gfp expression significantly increased when the fluorescence intensity was up to 62 000 at presence of 25 μg/mL 4-isopropylbenzoic acid for 12 h.This proved that PH 10-CuO has very good rigor and induced expression intensity.Meanwhile,in Constructing the gene editing plasmid for C.glutamicum with PH10-CuO regulating the expression of recET and cas12a,The accurate editing of target gene and insertion of foreign genes in C.glutamicum chromosome.[Conclusion]The inducible promoter PH10-CuO presented the advantages with high intensity and low leakage,which enables it as an useful element to regulate temporal expression of specific genes in C.glutamicum.
Corynebacterium glutamicuminducible promoterrepressor protein CymR4-isopropylbenzoic acid
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