Cloning and expression of E7 gene of human papillomavirus type 16 (HB 
strain

WU Xin-Xing; ZHAO Wen-Xian

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August 1998

WU Xin-Xing and ZHAO Wen-Xian. Cloning and expression of E7 gene of human papillomavirus type 16 (HB strain[J]. Virologica Sinica, 1998, 13(4).

Citation: WU Xin-Xing, ZHAO Wen-Xian. Cloning and expression of E7 gene of human papillomavirus type 16 (HB strain .VIROLOGICA SINICA, 1998, 13(4) : 326.

人乳头瘤病毒16型湖北株E_7基因的克隆和高效表达

伍欣星 ,
赵文先

1.
湖北医科大学病毒研究所

摘要

利用基因克隆技术，将ＨＰＶ１６湖北株完整的Ｅ７基因克隆到含乳糖操纵子的表达载体ｐＷＲ５９０１上，经限制性酶切分析获得重组质粒ｐＷＨＢＥ７。ｐＷＨＢＥ７转化大肠杆菌后表达产生分子量为７０ｋＤ的融合蛋白ｌａｃＥ７，该蛋白在免疫印迹实验中可被标准Ｅ７单抗识别。经ＩＰＴＧ诱导后，Ｅ７融合蛋白产量可达细菌总蛋白含量的３０％以上。利用ｌａｃＥ７蛋白在细菌胞浆中形成包含体的性质，简便地提取并纯化了该蛋白质。结果为从免疫学角度探讨ＨＰＶ１６与宫颈癌的关系以及ＨＰＶ疫苗的研制打下基础。
- 人乳头瘤病毒克隆表达融合蛋白包含体 HPVE7蛋白

Cloning and expression of E7 gene of human papillomavirus type 16 (HB strain

1.
Virus Research Institute．Hubei Medical University．Wuhan 430071

Abstract

HPV 16 (HB strain) E 7 gene was cloned into expression vector pWR590 1 by gene cloning technique. Recombinant plasmid pWHB E 7 was obstained by analysis of restrictional endonuclease. 70 kD HPV 16 (HB strain) E 7 fusion protein was expressed by E.coli JM 109 transfected with pWHB E 7. The recombinant protein could be recognized by standard E 7 monocloned antibody in Western blot. The production of E 7 recombinant protein was more than 30 percent of total E.coli protein by induction of IPTG. The E 7 fusion protein existing in inclusion body in E. coli could be conveniently extracted and purified.
- HPV Cloning expression Fusion protein Inclusion body HPV E 7 protein

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Cloning and expression of E7 gene of human papillomavirus type 16 (HB strain

1. Virus Research Institute．Hubei Medical University．Wuhan 430071

Abstract: HPV 16 (HB strain) E 7 gene was cloned into expression vector pWR590 1 by gene cloning technique. Recombinant plasmid pWHB E 7 was obstained by analysis of restrictional endonuclease. 70 kD HPV 16 (HB strain) E 7 fusion protein was expressed by E.coli JM 109 transfected with pWHB E 7. The recombinant protein could be recognized by standard E 7 monocloned antibody in Western blot. The production of E 7 recombinant protein was more than 30 percent of total E.coli protein by induction of IPTG. The E 7 fusion protein existing in inclusion body in E. coli could be conveniently extracted and purified.