Construction of HIV-1 B subtype pseudovirus system in Guangxi
HUANG Chunyuan
WANG Hong
LIANG Hao
YE Li
LIANG Bingyu
JIANG Junjun
CHEN Rongfeng
NING Chuanyi
LIAO Yanyan
YU Jun
HUANG Jiegang
Abstract：Objective To establish a pseudovirus system for phenotypic drug-resistance detection and provide a relatively cheap and easy method for drug-resistance testing. Methods EGFP gene was amplified from plasmid pSV-EGFP and then cloned to backbone plasmid pNL4-3.Luc. E-R-by double enzyme digestion;env gene was amplified from RNA isolated from HIV-1-infected persons and cloned to eukaryotic expression plasmid cells and EGFP or ENV expression. Pseudovirus was produced by co-transfection of two recombinant plasmids to 293t cells. Infection of pseudovirus was determined by co-cultured with TZM-b1 cells and immunofluorescent test. Results Two recombinant plasmids(mass ratio,pcDNA3.1-env:pNL4-3.EGFP.E-R-.=2:1)were co-transfected to 293t cells. Cultured supernatants containing pseudovirus were harvested at 48 h post-transfection. Fluorescence was observed in TZM-b1 cells after TZM-b1 cells were infected with pseudovirus at 48 h post-infection. Conclusion The recombinant pseudovirus carrying EGFP gene is constructed successfully and it could be used for phenotypic drug-resistance detection.
Keywords：HIV-1pseudovirusdrug-resistance detection
Publication Date：2018-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 1942-1946 )