疹病_____](期刊论文)

人疱疹病毒7型U14编码区原核表达载体的构建

http://www.100md.com 《青岛大学医学院院报》 2006年第1期

疹病,,],疱疹病毒7型,人；被膜蛋白pp85；原核表达,1材料和方法,2结果,3讨论,[参考文献]

     [摘要] 目的构建人疱疹病毒7型(HHV7)开放读码框(ORF) U14的原核表达载体，并且进行原核表达。方法 PCR技术扩增HHV7 ORF U14基因的主要抗原决定簇编码区(328～533氨基酸)，目的基因与原核表达载体pThioHis A分别双酶切后连接，1×TSS法转化宿主菌E.coli BL21，异丙基βD硫代半乳糖苷诱导融合蛋白表达。结果基因序列分析表明目的基因序列与HHV7标准毒株相应序列基本一致，SDSPAGE电泳可观察到相对分子质量为3.57万融合蛋白的表达，Western印迹证实pp85蛋白成功表达。结论 HHV7被膜蛋白pp85原核表达载体构建和表达成功，为进一步探讨该重组蛋白在HHV7特异性抗体检测中的应用提供实验基础。

    [关键词] 疱疹病毒7型,人；被膜蛋白pp85；原核表达

    CONSTRUCTION OF PROKARYOTIC EXPRESSION CLONE OF HUMAN HERPESVIRUS 7 U14 CODING REGIONSUN JIANPING，WANG XIAOFENG，WANG YUN，et al(Department of Microbiology，Qingdao University Medical College，Qingdao 266021,China)

    [ABSTRACT]ObjectiveTo construct the prokaryotic expression vector of tegument protein pp85 of human herpesvirus 7 and to express it. Methods The major antigen epitopes coding sequence (328—533 aa) of HHV7 ORF U14 gene was amplified by PCR and inserted into the prokaryotic expression vector pThioHis A by gene engineering technique; recombinant plasmid was transformed into E.coli BL21 and expressed by IPTG inducing. ResultsThe target gene was identical with that of HHV7 standard species，and the 35 700 fusion protein was found in SDSPAGE and proved by western blot. ConclusionRecombinant prokaryotic expression vector was gained, which is valuable for the further study on HHV7 specific antibody detection. ......

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