HPV11-E7与人干扰素α-2b融合表达质粒的构建及表达  

Construction and Expression of Plasmid Coexpressing Human Papillomavirus Type 11 E7 and Human IFNα-2b

作　　者：相文忠[1] 王飞[1] 李光富[2] 王新军[2] 王群[3] 刘丰[2] 张兆松[2] 毕志刚[1] 

机构地区：[1]南京医科大学第一附属医院皮肤科,210029 [2]南京医科大学分子生物研究所 [3]广东省人民医院皮肤科

出　　处：《中华皮肤科杂志》2005年第10期640-642,共3页Chinese Journal of Dermatology

基　　金：江苏省重点学科基金(135-03)

摘　　要：目的构建HPV11-E7与人干扰素a-2b(IFNa-2b)融合基因表达载体,并进行原核表达。方法通过连接肽将HPV11-E7与人IFNa-2b连接并克隆至原核表达载体pE7-32a,进行酶切鉴定及DNA序列分析。将重组质粒转染E．coli BL21,经IPTG诱导后对其表达产物进行SDS-PAGE和Westernblotting分析。结果测序结果表明将HPV11-E7和人IFNa-2b通过连接肽(G-G-S-G-S)_3连接并克隆至pET-32a,且其基因序列与设计完全一致。SDS-PAGE和Western blotting分析结果均显示在相对分子质量约50 000位置可见明显蛋白条带。结论 HPV11-E7与人IFNa-2b融合基因表达载体的成功构建及表达,为今后的相关实验研究奠定了基础。Objective To construct an expression plasmid of human papillomavirus type 11 E7 （HPV11-E7）/human IFNα-2b fusion gene,to express the fusion gene in E.coli BL21, and pave way for further immunological study, Methods The recombinant plasmid was introduced into E.coli BL21, then the expression product was analyzed by SDS-PAGE and Western blotting after induction with isopropy-β-D-thiogalactoside （IPTG）. Results The fusion gene of HPVll-E7 and human IFNα-2b was successfully cloned into pET-32a by a linker with the same sequence as we expected, The expressed fusion protein was confirmed by SDS-PAGE and Western blotting. Conclusions The successful construction of prokaryotic expression plasmid and expression of HPV11-E7/human IFNα-2b fusion gene enable further immunological study.

关 键 词：尖锐湿疣 人乳头瘤病毒 E7基因 IFNΑ-2B 融合基因 干扰素α-2b 融合表达质粒 BLOTTING Western 基因表达载体 

分 类 号：R752.53[医药卫生—皮肤病学与性病学]