HIV-1跨膜蛋白gp41螺旋结构的原核表达及功能研究

HIV-1跨膜蛋白gp41是HIV-1包膜与靶细胞膜的融合过程中的关键蛋白，是理想的HIV-1融合抑制剂靶点。为开展以gp41为靶点的抑制剂筛选，以HIV-1 B亚型病毒基因为模板，通过PCR、酶切、连接等方法构建得到gp41 5-helix与6-helix重组质粒，转入大肠杆菌BL21（DE3）进行表达，经变性和复性后亲和层析纯化蛋白。经SDS-PAGE鉴定，纯化后蛋白纯度较高。本研究还通过非变性凝胶电泳证明gp41 5-helix与C-多肽衍生物T-20存在相互作用，为下一步药物筛选模型的建立奠定了基础。

Prokaryotic expression and functional study of HIV-1 envelope glycoprotein gp41 helical bundle

HIV-1 envelope glycoprotein gp41, which is a hopeful target for HIV-1 fusion inhibitors, plays a critical role in the fusion of viral and cellular membranes. In order to build up the screening assay of HIV-1 fusion inhibitors targeting gp41, HIV-1 gp41 5-helix and 6-helix were expressed in prokaryotic cells. Gp41 5-helix and 6-helix recombined plasmids were constructed by using PCR, enzyme digestion and ligation taking the clade B HIV-1 genome as a template. The plasmid was transferred into E. coli BL21（DE3）and then induced by IPTG. The expressed protein was purified by affinity chromatography after denaturation and renaturation. The SDS-PAGE analysis was used during expression and purification. Native-PAGE was used to identify the interaction between gp41 5-helix and T-20. It will be helpful to build up the screening assay of HIV-1 fusion inhibitors targeting gp41.