Prokaryotic Expression, Purification and Identification of Human Papillomavirus Type 58 E6E7 Fusion Protein

Objective: To construct pET-42a(+)-HPV58E6E7 prokaryotic expression plasmid and to express and purify human papillomavirus type 58(HPV58) E6E7 fusion protein.Methods:The HPV58 E6E7 fusion gene were amplified by PCR and cloned into pET-42a(+).The recombinant plasmids were successfully introduced into E.co li BL21 and were induced by IPTG.SDS-PAGE and Western blot analysis were used to detect the fusion protein.Results:The recombinant plasmid of pET-42a(+)-HPV58E6E7 was identified and confirmed with enzyme diges tion and sequencing.HPV58E6E7 fusion protein was expressed and purified sucessfully.SDS-PAGE and Western blot showed that the fusion protein was correct and effective.Conclusion: pET-42a(+)-HPV58E6E7 prokaryotic expression plasmid was constructed successfully.High-level expression of HPV58E6E7 fusion protein was achieved and purified.The fusion protein can be used as antigen of immunological test to HPV58 therapeutic vaccine.