摘要：目的用细菌／杆状病毒（BactoBac）表达系统在昆虫细胞Sf9中表达HIV-2外膜糖蛋白gp105和跨膜糖蛋白gp36，为研制艾滋病疫苗和诊断试剂奠定基础。方法分别将HIV-2外膜蛋白gp105和跨膜蛋白gp36全基因序列克隆到杆状病毒转座载体pFastBacHTa和pFastBacHTb中多角体启动子下游，构建成重组转座载体pFastBacHTa-gp105和pFastBacHTb-gp36，利用细菌／杆状病毒（BactoBac）表达系统筛选重组杆状病毒，并在昆虫细胞Sf9中表达HIV-2的gp105和gp36。结果SDS-PAGE分析结果表明，gp105基因表达产物为一66000u糖蛋白，gp36基因则表达一41000u糖蛋白，与天然产物一致。Westernblot结果显示：两者均具有抗原特异性。结论gp105和gp36在昆虫细胞中得到了高效表达，并均被糖基化；gp105接近天然产物，gp36与天然产物一致。

分类号：R392文献标识码：A

文章编号：1000-8861（2001）01-0013-04

Expression of external glycoprotein gp105 and transmembrane glycoprotein gp36 of human immunodeficiency virus type 2 in insect cell by Bac-to-Bac system

ZHANG Ying-jiu（College of Life Science， Jilin University， Changchun 130023，China）

JIN Ning-yi（Genetic Engineering Laboratory， the Quartermaster University of PLA， Changchun 130062，China）

KIMSun-yong（Genetic Engineering institute of Seoul University）

WANG Hong-wei（Genetic Engineering Laboratory， the Quartermaster University of PLA， Changchun 130062，China）

SHEN Jia-cong（College of Life Science， Jilin University， Changchun 130023，China）

Abstract：ObjectiveTo develop effective vaccine and diagnostic agents of AIDS， the external glycoprotein gp105 and transmembrane glycoprotein gp36 of human immunodeficiency virus type 2 were expressed in insect cells Spodoptera frugiperda (Sf9) using baculovirus expression system ： Bac-to-Bac system. MethodsThe recombinant transfer vector pFast Bac HTa-gp105 and pFast Bac HTb-gp36 were constructed by cloning HIV-2 gp105 gene and gp36 gene into the end of polyhedrin promoter of baculovirus transfer vector pFast Bac HTa and pFast Bac HTb respectively. HIV-2 external glycoprotein gp105 and transmembrane glycoprotein gp36 were expressed in insect cells Spodoptera frugiperda (Sf9) by transposing these two foreign genes to shuttle vector bacmid respectively using Bac-to-Bac system. ResultsThe resulting products determined by SDS-PAGE showed that recombinant baculovirusescontaining gp105 gene expressed a 66 kDa glycoprotein， whereas those contai-ning gp36 gene generated a 41 000u glycoprotein， the apparent molecular weight of which is equivalent to that of natural gp36. Western blot indicated that both gp105 and gp36 showed antigenic specificity. Conclusionsgp105 and gp36 has been expressed in insect cells Spodoptera frugiperda (Sf9) using baculovirus expression system.

Keywords：HIV-2；gp105；gp36；baculovirus；Spodoptera frugiperda (Sf9)基金项目：国家杰出青年基金资助项目（39825119）

作者简介：张应玖（1962－），女，吉林长春市人，副教授，博士，主要从事蛋白质结构与功能的研究。Tel：（0431）8922331-3762； e-mail：zyingjiu＠public.cc.jl.cn参考文献：

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收稿日期：2000年8月29日

修稿日期：2000年10月16日

出版日期：2001年1月15日