重组HIV表面抗原gp120的表达纯化及免疫学鉴定

为研制具有流行特点的HIV血清学诊断试剂，采用pET系统表达HIV-1表面糖蛋白gp120。研究发现，全长的gp120在E.coli中不能有效表达；N端半长的gp120可以表达，但表达量很低；仅保留N端1/3的gp120(包含gp120V1/V2抗原决定簇)有效表达，表达蛋白占菌体总蛋白的18%；Westernblot显示较好的反应原性；通过金属螯合层析，产物得到完全纯化。在这些结果的基础上，我们表达了流行株的gp120片段，为探索gp120在大肠杆菌的高效表达，建立针对中国人群的HIV血清学诊断系统奠定基础。

High Expression, Purification and Immuno-indentification of Recombinant HIV Surface Antigen gp120

Different-length fragments of the HIV trans-membrane antigen (gp120) gene were expressed in Escherichia coli, using T7 expression system. SDS-PAGE stained by Coomassic Brilliant Blue showed no expression of full-length gp120 and poor expression of half-length gp120 fragments from the N-terminal, but high expression of 1/3-lgenth gp120 gene fragments from the N-terminal (including V1/V2 epitopes), more than 18% of total bacterial protein. Western blot showed fairly good reactivity to serum from HIV-infected individual. On this basis, we expressed the corresponding tp 120 fragments of the HIV strains epidemic in China. This study laid a solid foundation for exploration of high expression of gp 120 in E.coli and development of HIV serological diagnostic system for Chinese people.