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.
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 target 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.
Lee
Myung Kyu
MK
Proteome Research Laboratory, Peptide Engineering National Research Laboratory, Korea Research Institute of Bioscience and Biotechnology, P.O. Box 115, Yusong, 305-600, Taejon, South Korea. mklee@mail.kribb.re.kr
Seo
Jeong Kon
JK
Kim
Hee Kyung
HK
Cho
Ju Hyun
JH
Poo
Haryoung
H
Kim
Kil Lyong
KL
D002460:Cell Line; D015686:Gene Products, env / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015752:Genes, env / Q000235:genetics*; D005822:Genetic Vectors; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000472:pathogenicity; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics*; D009500:Neutralization Tests; D010957:Plasmids; D011994:Recombinant Proteins; D014162:Transfection; D014771:Virion / Q000235:genetics
D015686:Gene Products, env; D011994:Recombinant Proteins