The Production And Identification Of Human Papillomavirus Type58Produced In Eschrichia Coli

In the world,cervical cancer is one of the common gynecologic malignancies,anincrease of about500,000every year, ranking second in the incidence of cancer inwomen, following breast cancer.Human Papillomavirus (HPV)is a member of the pa-poviridae family of viruses, non-enveloped DNA, the main cause of human skin andmucous membranes of proliferative lesions, highly relevant to humanpathogens. HPVis the main factor in cervical cancer,included the expression of almost all cervicalcancer cells in both HPV DNA and virus-transformed. A large number of molecularepidemiology and molecular biology studies have demonstrated that high-risk HPVinduced cervical cancer. HPV58type is the third most important type of cervical can-cer in women,following the type16and18. Since HPV have species-specific differen-tiation and proliferation strictly due to host cell,it is difficult to culture in vitro,the tra-ditional inactivated and live attenuated vaccines being limted.The focus on the studyis to turn to the genetically engineered vaccine. VLP L1protein is the major forma-tion of HPV vaccine, in the development of HPV VLP vaccine and how to completethe preparation of simple and efficient structure, strong immunogenic VLP is the keyto all problems.In this study,the HPV-58-L1gene was expressed in Escherichia coli as a fusionprotein with a glutathione-S-transferase tag.The conditions of expression,such asIPTG concentration,induced time and induced temperature were optimized.Because ofthe complex space structure and others factors,the HPV58L1protein is mainly inclu-sion body.The refolded HPV58L1protein is obtained,takeing measures is gradientdialysis,after denaturing in urea.Transfering chaperone，and selecting the optimalcondition we get the active protein in the supernatant,as at the temperature of15℃cold-induced expression of the maximum amount of soluble expression. Through thecolumn purification, western blotting, the results showed that the expression ofHPV58L1protein could react with monoclonal HPV16subtypes We analyze the im-mugensity of HPV58L1protein,providing the foundation developing the new casevaccines.