人乳头瘤病毒16型湖北株E_7基因的克隆和高效表达  

Cloning and Expression of E 7 Gene of Human Papillomavirus Type 16 (HB Strain)

作　　者：伍欣星[1] 赵文先[1] 

机构地区：[1]湖北医科大学病毒研究所

出　　处：《中国病毒学》1998年第4期322-326,共5页Virologica Sinica

基　　金：湖北省科委"八五"重点攻关课题

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。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.

关 键 词：人乳头瘤病毒 克隆表达 融合蛋白 HPVE7蛋白 

分 类 号：R373.9[医药卫生—病原生物学] Q78[医药卫生—基础医学]