利用基因工程技术制备HPV58E7蛋白  

The preparation of HPV58E7 protein by GST-tagged prokaryotic expression system

作　　者：王德普[1] 霍正浩[1] 张晨华[2] 闫小飞[2] 

机构地区：[1]宁夏医科大学医学遗传学教研室,宁夏银川750004 [2]西安交通大学生命学院癌症研究所,陕西西安710061

出　　处：《西北大学学报（自然科学版）》2011年第2期265-267,共3页Journal of Northwest University（Natural Science Edition）

基　　金：教育部博士点基金资助项目(20070698097)

摘　　要：目的利用基因工程技术制备HPV58E7蛋白。方法设计E7基因片段特异性PCR引物,以HPV58全基因组为模板,经PCR扩增E7基因片段,利用pGEX-4T3表达质粒,构建pGEX-4T3/HPV58E7重组体,转化BL21宿主细胞,IPTG诱导表达带有GST标签的可溶性融合蛋白。经谷胱甘肽凝胶柱亲和层析纯化,凝血酶切GST标签。结果 SDS-PAGE分析显示在分子量约为40KD处有新生蛋白条带,经凝血酶消化融合蛋白,所获纯化HPV58E7蛋白与预期理论值相吻合。结论利用带有GST标签的原核表达系统及其相应纯化策略,获得HPV58E7蛋白的高效制备。Aim To prepare HPV58E7 protein by genetic engineering technique.Methods Using HPV58 genome as the template,HPV58E7 fragment was amplified by polymerase chain reaction,the pGEX-4T3/HPV58E7 recombinant plasmid was constructed by inserting the HPV58E7 fragment into pGEX-4T3,the GST-tagged expression plasmid and BL21 strain as the host cell.After induction by IPTG,soluble GST-E7 protein was expressed and purified with glutathione sepharose 4B affinity chromatograph.Results The expressed GST-HPV58E7 protein with 40KD was revealed by SDS-PAGE and purified HPV58E7 without GST-tag was accomplished by digesting with thrombin at the thrombin cleavage site.Conclusion HPV58E7 protein was efficiently produced with GST-tagged expression system.

关 键 词：GST-HPV58E7融合蛋白 HPV58E7蛋白 人乳头瘤病毒58型 

分 类 号：Q812[生物学—生物工程]