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来自重组杆状病毒感染昆虫细胞的HIV-2 gag前体蛋白表达及病毒样gag颗粒分泌

Expression of gag precursor protein and secretion of virus-like gag particles of HIV-2 from recombinant baculovirus-infected insect cells.

作者信息

Luo L, Li Y, Kang C Y

机构信息

Department of Microbiology and Immunology, University of Ottawa, Faculty of Medicine, Ontario, Canada.

出版信息

Virology. 1990 Dec;179(2):874-80. doi: 10.1016/0042-6822(90)90159-o.

Abstract

A recombinant baculovirus carrying the gag gene but lacking the protease coding sequences of human immunodeficiency virus type 2 (HIV-2) has been constructed. When this recombinant baculovirus is used to infect insect cells, a high level of gag precursor protein, gag pr41, is expressed. Electron microscopy showed that the majority of gag pr41 was budding through the plasma membrane and being released into the culture medium in spherical virus-like particles with a diameter of approximately 100 nm. Metabolic labeling demonstrates that gag pr41 is myristylated. Our results demonstrated that HIV-2 gag pr41 can be assembled into virus-like particles in the absence of other HIV proteins. Rabbits immunized with purified gag pr41 particles produced high-titer antibody and Western blot analysis showed that anti-gag pr41 rabbit sera recognize p17, p24, and p55 gag proteins of HIV-1. These results show that gag pr41 particles are highly immunogenic and that gag proteins of HIV-1 and HIV-2 have similar antigenic epitopes.

摘要

已构建出一种携带2型人类免疫缺陷病毒(HIV-2)gag基因但缺失蛋白酶编码序列的重组杆状病毒。当用这种重组杆状病毒感染昆虫细胞时,会表达出高水平的gag前体蛋白gag pr41。电子显微镜显示,大多数gag pr41通过质膜出芽,并以直径约100 nm的球形病毒样颗粒形式释放到培养基中。代谢标记表明gag pr41被肉豆蔻酰化。我们的结果表明,在没有其他HIV蛋白的情况下,HIV-2 gag pr41可以组装成病毒样颗粒。用纯化的gag pr41颗粒免疫的兔子产生了高滴度抗体,蛋白质印迹分析表明,抗gag pr41兔血清可识别HIV-1的p17、p24和p55 gag蛋白。这些结果表明,gag pr41颗粒具有高度免疫原性,且HIV-1和HIV-2的gag蛋白具有相似的抗原表位。

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