title: [Construction of prokaryotic expression vector of HPV16E6 gene and its expression].
abstract: Prokaryotic expression vector of mouse HPV16E6 gene was constructed. A pair of primers were designed according to the digestion sites in plasmid pGEX-KG and the HPV16E6 gene sequence published by GenBank. The DNA fragment of 321bp was amplified by PCR from the HPV recombinant plasmid with HPV16E6 gene, then cloned into pGEX-KG and transformed into the host E. coli strain JM109. The fragment was conformed to the original sequence, which indicated that fusion expression vector pGEX-KG-HPV16E6 was constructed. The pGEX-KG-HPV16E6 plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37 degrees C, the expression product of HPV16E6 gene was identified by SDS-PAGE and Western blot. HPV16E6 fusion protein had been expressed successfully in the form of inclusion bodies, the molecular weight of fusion protein being 38 kD. Meanwhile, the optimum condition of HPV16E6 fusion protein expression was induced with 1.0 mmol/L IPTG for 4h. The fusion protein reacted specifically with the antibodies against HPV16E6. HPV16E6 gene was successfully expressed in E. coli, which could be used as a basis for preparing HPV16E6 vaccine in human.
authors.affiliation: School of Chemistry and Bioengineering, Chongqing University of Technology, Chongqing 400050, China.
authors.forename: Fangyi
authors.initials: F
authors.lastname: Peng
authors.forename: Hairong
authors.initials: H
authors.lastname: Jiang
authors.forename: Fangliang
authors.initials: F
authors.lastname: Peng
authors.forename: Weibing
authors.initials: W
authors.lastname: Zhao
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Lin
authors.forename: Yuanxiang
authors.initials: Y
authors.lastname: Chen
authors.forename: Shengzhen
authors.initials: S
authors.lastname: Chen
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D014765:Viral Vaccines / Q000276:immunology
chemical_list: C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014765:Viral Vaccines; D005982:Glutathione Transferase