Cloning and expression of HPV16 E7 gene

Objective:To study the expression and activity of the gene E7 protein of human papillomavirus in the expression system of the prokaryon and built an experimental foundation for preparing the E7 recombinant protein. Methods:HPV16E7 gene amplified by PCR was cloned into the expression vector pET32a to form recombinant plasmid pET32a/ HPV 16E7,then transformed into E. coli BL21. The pET32a/ HPV16 E7 fusion protein was induced by IPTG and purified with Ni-NTA gel chromatography column. The protein was verified with SDS-PAGE and Western blot. Results:The recombinan plasmid was identified and confirmed with double enzymes digestion and sequencing. It was suggested that the recombinant E7 protein was expressed with SDS-PAGE and western blot, and a molecular weight was about 30×103. The fusion protein was about 16% of the total bacterial protein. Conclusions:Highly expressed E7 protein of HPV16 is obtained.