A vector system for introducing foreign HIV-1 env genes and pseudotyping of MuLV particles with the recombinant HIV-1 envelope proteins for anti-HIV-1 assay

被引:69
作者
Lee, MK
Seo, JK
Kim, HK
Cho, JH
Poo, H
Kim, KL
机构
[1] Korea Res Inst Biosci & Biotechnol, Proteome Res Lab, Peptide Engn Natl Res Lab, Taejon 305600, South Korea
[2] Sungkyunkwan Univ, Dept Sci Biol, Suwon 440746, Kyunggi Do, South Korea
关键词
HIV-1 env cloning vector; HIV-1/MuLV pseudotypes; anti-HIV-1; assay;
D O I
10.1016/S0166-3542(01)00196-6
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
We attempted to incorporate the HIV-1 envelope proteins derived from various HIV-1 strains into MuLV particles for developing a rapid and safe anti-HIV-1 screening system. In a previous study, only HIV-1 envelope protein lacking cytoplasmic 144 amino acids has been reported to be able to incorporate into MuLV particles. We designed and constructed a vector, pcKCX, expressing the envelope glycoprotein with cytoplasmic truncation by introducing the partial foreign HIV-1 env gene corresponding to the ectodomain of its envelope protein. Three HIV-1 env genes of AD8, BaL or 89.6 strains were cloned, and the HIV-1/MuLV pseudotypes were generated in the transfected TELCeB6 cells with all the cloned plasmids. The pseudotypes displayed host specificity depending on their original HIV-1 strains and their infection to the tar-et cells was inhibited by treatment of a potent anti-HIV-1 peptide C34. A stable cell clone against the HIV-1(BaL) strain was found to express the R5 tropic envelope glycoprotein on the cell surface and to produce continuously HIV-1(BaL)/MuLV pseudotypes. These results suggested that the vector system is useful for cloning of various foreign HIV-1 env genes and the recombinant envelope glycoproteins effectively incorporate into MuLV particles. The HIV-1/MuLV pseudotypes may be useful for anti-HIV-1 assay. V 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:99 / 111
页数:13
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