Construction of HIV-1 B subtype pseudovirus system in Guangxi
10.3969/j.issn.1006-5725.2018.12.006
- VernacularTitle:广西HIV-1 B亚型假病毒的构建
- Author:
Chunyuan HUANG
1
;
Hong WANG
;
Hao LIANG
;
Li YE
;
Bingyu LIANG
;
Junjun JIANG
;
Rongfeng CHEN
;
Chuanyi NING
;
Yanyan LIAO
;
Jun YU
;
Jiegang HUANG
Author Information
1. 广西医科大学公共卫生学院&广西艾滋病防治研究重点实验室
- Keywords:
HIV-1;
pseudovirus;
drug-resistance detection
- From:
The Journal of Practical Medicine
2018;34(12):1942-1946
- CountryChina
- Language:Chinese
-
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.
