CAJ | 학술논문

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gp120 V1/V2抗原决定簇)有效表达，表达蛋白占菌体总蛋白的18%；Western blot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。
위연제구유류행특점적HIV혈청학진단시제，채용pET계통표체HIV-1표면당단백gp120。연구발현，전장적gp120재E.coli중불능유효표체；N단반장적gp120가이표체，단표체량흔저；부보류N단1/3적gp120(포함gp120 V1/V2항원결정족)유효표체，표체단백점균체총단백적18%；Western blot현시교호적반응원성；통과금속오합층석，산물득도완전순화。재저사결과적기출상，아문표체료류행주적gp120편단，위탐색gp120재대장간균적고효표체，건립침대중국인군적HIV혈청학진단계통전정기출。
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.