Construction and Functional Analysis of Prokaryotic and Eukaryotic Dual Expression Vectors
ZHU Minghui, LI Xiaojing, WANG Haomin, QIAO Kexin, ZHU Jiayi, ZHAO Chunpeng*, WANG Xiaoyin*
This study was to construct a dual expression vector that could simultaneously drive the expression of recombinant protein in eukaryotic and prokaryotic expression system. The T7 promoter and T7 terminator gene of prokaryotic were cloned into the downstream of CMV promoter and upstream of neomycin resistance gene of eukaryotic expression vector pIRES-EGFP to generate the dual expression vector pIRES-CMV/T7-EGFP, and further the human transferrin gene was replaced with eGFP (enhanced green fluorescent protein) gene to generate pIRES-CMV/T7-HTrf vector. Subsequently, the recombination vector was transfected into CHO (Chinese hamster ovary) cells and transformed into Escherichia coli BL21. The results showed that the eGFP and HTrf gene could be successfully expressed in both CHO cells and E. coli BL21 by fluorescence microscopy, flow cytometry and Western blot. Taken together, this study has successfully constructed a dual expression vector that can express the recombinant protein in both prokaryotic and eukaryotic expression systems.
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