pGEX-4T-1/HSV2-gG重组质粒的构建及表达  

Construction and expression of the recombinant plasmid of pGEX-4T-1/HSV2-gG

作　　者：戍桂明 杨慧兰[2] 成桂明[1] 

机构地区：[1]武警广东省总队医院皮肤性病科,广东广州510507 [2]广州军区广州总医院皮肤性病科,广东广州510010

出　　处：《中国现代医学杂志》2009年第9期1342-1345,共4页China Journal of Modern Medicine

摘　　要：目的通过基因工程技术,构建含有目的基因的重组质粒(pGEX-4T-1/HSV2-gG)。方法用PCR技术扩增HSV2-gG基因的免疫优势表位片断;将PCR产物与质粒表达载体pGEX-4T-1连接;然后鉴定含有目的基因的重组表达载体;并转化大肠杆菌DH5α,诱导表达目的蛋白;以免疫印迹法检测表达的目的蛋白。结果PCR法扩增出1242bp的DNA片断;通过PCR法及测序法证实重组质粒中含有1242bp的DNA片断。SDS-PAGE和免疫印迹法证实重组蛋白在大肠杆菌中的表达。结论重组质粒HSV2-gG-pGEX-4T-1的成功构建及表达,为制备HSV-2血清诊断试剂奠定了基础。[Objective] To construct and express the recombinant plasmid of pGEX-4T-1/HSV2-gG. [Methods] The target gene was amplified by polymerase chain reaction （PCR）. The PCR products were ligated into pGEX- 4T-1 expression vector. After identification, the recombinant expression vector was transferred into E.coli JM109 for expression. Finally, recombinant protein of FgG-2 （rFgG-2） was detected by Western Blot （WB）. [Results] A 1242 bp DNA fragment was obtained with PCR and then confirmed in recombinant vector by PCR and sequencing. Ex- pression of target protein was confirmed by WB with anti-gG monoclonal antibody. [Conclusions] The recombinant plasmid of pGEX-4T-1/HSV2-gG has been successfully constructed and expressed in E.coli, which might be used for the development of serum diagnostics assay kits for HSV-2 infection.

关 键 词：HSV-2-gG 重组质粒 基因表达 

分 类 号：R782[医药卫生—口腔医学]