重组HIV表面抗原gp120的表达纯化及免疫学鉴定

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

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gp12 0。研究发现 ,全长的 gp12 0在E .coli中不能有效表达 ;N端半长的gp12 0可以表达 ,但表达量很低 ;仅保留N端 1/3的 gp12 0 (包含gp12 0V1/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 gp12 0片段 ,为探索gp12 0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。

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.