Prokaryotic cloning and expression of HPV16 E7 gene

Objective:To explore the cloning and expression of HPV16 E7 protein from laryngeal carcinoma. Methods: HPV16 E7 gene was amplified by PCR. The amplified fragment was inserted into the plasmid pET28a (+) digested with BamHⅠand Hind Ⅲ. The recombinant plasmid pET28/E7 was transformed into E.coli JM109 which was selected with ampicillin. HPV16 E7 recombinant protein expression in the E.coli BL21(DE3) was identified by SDS-PAGE and Western blot. Results: The prokaryotic recombinant plasmid pET28/E7 was successfully constructed. The BL21(DE3) transformed recombinant plasmid pET28/E7 had expressed HPV16 E7 recombinant protein effectively. Conclusion:The construction of the prokaryotic recombinant plasmid pET28/E7 and the successful expression of the recombinant protein HPV16 E7 pave way for the profound research of the biological properties and the transformational mechanism of the HPV16 E7 protein on the specific cells.