中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (10): 640-642.

• 论著 • 上一篇    下一篇

HPV11-E7与人干扰素α-2b融合表达质粒的构建及表达

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   

  1. 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
  • 收稿日期:2004-10-07 发布日期:2005-10-15
  • 通讯作者: 毕志刚,E-mail:eltonbi@21cn.com E-mail:eltonbi@21cn.com
  • 基金资助:
    江苏省重点学科基金(135-03)

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

XIANG Wen-zhong1, WANG Fei1, LI Guang-Ju2, WANG Xin-jun2, WANG Qun3, LIU Feng2, ZHA NG Zhao-song2, BI Zhi-gang1   

  1. Department of Dermatovenerology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
  • Received:2004-10-07 Published:2005-10-15

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Western blotting分析。结果 测序结果表明将HPV11-E7和人IFNα-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50000位置可见明显蛋白条带。结论 HPV11-E7与人IFNα-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。

关键词: 尖锐湿疣, 人乳头瘤病毒, E7基因, IFNα-2b, 融合基因

Abstract: Objective To construct an expression plasmid of human papillomavirus type 11 E7 (HPV11-E7)/hurnan 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 HPV11-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.

Key words: Condyloma accuminatum, Human papillomavirus, E7 gene, IFNα-2b, Fusion gene