人疱疹病毒7型U14编码区原核表达载体的构建  

CONSTRUCTION OF PROKARYOTIC EXPRESSION CLONE OF HUMAN HERPESVIRUS 7 U14 CODING REGION

作　　者：孙坚萍[1] 王笑峰[1] 王云[1] 罗兵[1] 

机构地区：[1]青岛大学医学院微生物学教研室,山东青岛266021

出　　处：《青岛大学医学院学报》2006年第1期47-49,52,共4页Acta Academiae Medicinae Qingdao Universitatis

基　　金：青岛市科技局资助项目(2001KNS-2E-27-1)

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-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。Objective To 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 HHV70RF 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. coil BL21 and expressed by IPTG inducing. Results The 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. Conclusion Recombinant prokaryotic expression vector was gained, which is valuable for the further study on HHV-7 specific antibody detection.

关 键 词：疱疹病毒-7型 人 被膜蛋白pp85 原核表达 

分 类 号：R373[医药卫生—病原生物学]