Cloning and expression of E6 gene of human papillomavirus type 58

    ZHANG Ju, CHEN Zhongª²Can, BAI Yuª²Jie, GAO Yanª²E, HE Yuª²Xian, YAN Xiaoª²Jun

    1Institute of Gene Diagnostics, Fourth Military Medical University, Xi¡¯an 710033, China, 2Department of Obstetrics and Genecology, Second Hospital, Xi¡¯an Jiaotong University, Xi¡¯an 710004, China

    ¡¾Abstract¡¿ AIM: To obtain human papillomavirus type (HPV) 58 E6 gene and express it in vitro. METHODS: HPV58 E6 gene was amplified from DNA of a cervical cancer patient by PCR, and inserted into pGEMª²T Easy vector. HPV 58 E6 gene was digested from the recombinant plasmid HPV 58 E6ª²pGEMª²T, and ligated into lined prokaryotic expression vector pRSETª²A with SacI/Hind¢ó, and then expression of 58E6 protein was induced by IPTG. RESULTS: Fullª²length HPV58 E6 gene was successfully amplified from DNA of a cervical cancer patient by PCR. SDSª²PAGE analysis showed that His6ª²HPV58 E6 fusion protein of Mr 24 000 was expressed after induction of IPTG. The quantity of fusion protein was 10% of total bacterial proteins. CONCLUSION: HPV58 E6 gene is successfully cloned and effectively expressed in E. coli. ......