Expression and Purification of GST-HPV58 E6 Fusion Protein

Objective To clone and in vitro express HPV58 E6,and provide a good foundation for further study on the mechanism of carcinogenesis and preparing prevention vaccine.Methods Using PCR and gene recombinant,HPV58 E6 fragment was amplified from genome of HPV58 and inserted into the prokaryotic expression vector pGEX-4T3 with tag GST(glutathione sulfurtransferase).The recombinant plasmid was induced by IPTG to express GST-HPV58 E6 fusion protein.Results The recombinant plasmid was digested with restriction endonuclease BamHⅠ,XhoⅠ and analyzed by gene sequencing,which result indicated that HPV58 E6 gene was successfully inserted into pGEX-4T3 vector,and effectively expressed GST-HPV58 E6 fusion protein in E.coli DH5α.Furthermore,we obtained the purified the GST-HPV58 E6 fusion protein with Glutathione Sepharose 4B.Conclusion Prokaryotic expression vector pGEX-4T3/HPV58 E6 has been constructed successfully,and expressed GST-HPV58 E6 fusion protein,which will provide foundation for further study on the mechanism of carcinogenesis.