Recombinant Expression Of Truncated Human Papilloma Virus Subtype 58 L1 Protein With Molecular Chaperones In E.coli

Human papilloma virus is a kind of double stranded DNA virus,which has many subtypes.According to the difference of the infection,it can be divided into low risk type and high risk type.It causes common warts,flat warts,genital cancer,prostate cancer,oral cancer,cervical cancer,and other diseases through the skin and mucous membranes of the human body.In this paper,the study of the HPV 58 subtype is high-risk,and cervical cancer and cervical intraepithelial neoplasia is closely related to the occurrence of the infection of the HPV 58 subtype.Among women with cervical lesions in Asia,the rate of HPV 58 infection is high.And it is a serious threat to the health of the majority of women.However,domestic HPV vaccine research is relatively backward,there is no mature vaccine products,only the GSK two HPV(HPV16/18)vaccine in China.China is a high incidence of cervical cancer,the urgent need for independent research and development of effective HPV58 vaccine.The aim of this study is to develop an efficient and inexpensive HPV 58 L1 protein expression system,which lays the foundation for further preparation of HPV 58 preventive vaccine.In this study,the plasmid containing the molecular chaperone was transferred into the BL21 E.coli competent cell.And then the BL21 E.coli containing the molecular chaperone pTf16 was prepared into the state of perception.Then,the plasmid containing the target protein was transferred into the newly prepared competent cells.It was confirmed that the recombinant protein GST-HPV58 L1 and the molecular chaperone pTf16 could be expressed by the preliminary induction.The results showed that molecular chaperones could significantly promote the soluble expression of the target protein.By screening the concentration of IPTG,induction temperature,the concentration of L-Arabinose and the induction time of L-Arabinose,the optimal expression conditions were obtained.The ideal expression conditions of the target protein was L-Arabinose of 2.0 mg/ml,induced 20 min,after that added IPTG of 1.0 mmol/L,18 degrees Celsius to induce expression of 24 h.The expressed protein was purified by GST affinity chromatography column and cleaved by Protease Prescission.The cleaved protein was purified by GST affinity chromatography column.The results showed that the expression of the truncated HPV 58 L1 protein was successful,and the specific reaction bands appeared at 50 KD,and the purity of which was more than 90%.Kunming mice were immunized with the truncated HPV 58 L1 protein.The titer of antiserum was detected by ELISA.The results showed that there were some differences in the level of immune response among different mice.Both the high dose group(40 ?g/each mice)or the low dose group(20 ?g/each mice)could produce specific antibodies.Compared with PBS control group,there was significant difference.The preliminary results showed that the protein had good immunogenicity,which laid the foundation for the development of the vaccine.