中文摘要：

目的：表达和纯化人TRIM5α嵌合体蛋白[TRIM5αH（R328-332）],并探讨该蛋白和HIV-1gag间的相互作用。方法：将构建的TRIM5α嵌合体pET28aTRIM5αH（R328-332）,转化大肠杆菌BL21（DE3）,获得重组表达质粒pETTRIM5αH（R328-332）,30℃下经IPTG诱导,该融合蛋白在大肠杆菌BL21（DE3）中表达。获得的重组的蛋白再经过纯化,SDS-PAGE分析重组蛋白,并用免疫共沉淀技术和ELISA等检测重组蛋白与HIV-1gag间的相互作用。结果：构建的重组质粒在大肠杆菌中获得表达,重组TRIM5αH（R328-332）蛋白经纯化复性后,通过免疫共沉淀和ELISA等技术,证明TRIM5αH（R328-332）蛋白能够与HIV-1gag间的相互作用。结论：在大肠杆菌表达系统中成功表达了重组TRIM5αH（R328-332）蛋白,并且证实其在体外与HIV-1gag有结合作用。

英文摘要：

Objective： To express and purify the TRIM5α chimaera[ TRIM5α H（R328-332） ] protein and to explore the interaction between the TRIM5α H （R328- 332）and HIV-lgag. Methods： The plasmid pET28aTRIM5α H（R328-332） was transformed to E. coli BL21 （DE3） strain , and the expression of TRIM5α H（R328-332） protein was induced by IFFG, purified with Ni^2＋ chromatography. The expression and purification of TRIM5α H（R328-332） were analyzed by SDS-PAGE and Western blot,and the interaction between TRIM5α H （R328-332） and HIV-1 gag was detected by co-immunoprecipitation, His pull-down and ELISA. Results ： The recombinant plasmid pET28aTRIM5α H （R328-332 ） was successfully expressed in E. coll. The results showed that the purified full length TRIM5α H（R328-332） interacted with HIV-lgag protein. Conclusion： The human TRIM5α chimaera was expressed successfully in vitro, and the study demonstrates that the human TRIM5α chimaera interacts with HIV-1 gag in vitro.