Construction and characterization of an infectious clone of an HIV-1 CRF01_AE isolate from China
10.3760/cma.j.cn112866-20250731-00166
- VernacularTitle:我国HIV-1 CRF01_AE流行毒株感染性克隆的构建与病毒表型鉴定
- Author:
Jingwan HAN
1
;
Dijing JIA
1
;
Shuai CHANG
1
;
Hanping LI
1
;
Yongjian LIU
1
;
Lei JIA
1
;
Xiaolin WANG
1
;
Bohan ZHANG
1
;
Jingyun LI
1
;
Lin LI
1
Author Information
1. 中国人民解放军军事科学院军事医学研究院 病原微生物生物安全全国重点实验室，北京 100071
- Publication Type:Journal Article
- Keywords:
Human immunodeficiency virus（HIV）;
Infectious clone;
Full-length genome;
Phenotype
- From:
Chinese Journal of Experimental and Clinical Virology
2025;39(5):556-564
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct an infectious clone of a Chinese HIV-1 CRF01_AE epidemic strain with strong replication capacity，and comprehensively identify its viral phenotype and replication capacity.Methods:Using the CRF01_AE clinical isolate GX2005002，which was previously isolated from whole blood of an HIV-1-infected individual in China by our laboratory，as the parental strain，the full-length genome of the virus（9.7 kb）was divided into 5' half fragment（5.1 kb）and 3' half fragment（4.6 kb）for amplification. The proviral DNA was used as a template to amplify the virus genome，which was then ligated into the eukaryotic expression vector pEASY-T1. The consistency of its sequence with the parental strain sequence was identified through full-length genome sequencing and phylogenetic analysis. The replication capacity，syncytium inducibility，and cell tropism were experimentally identified to determine its phenotypic consistency with the parental strain.Results:An infectious clone of the CRF01_AE strain was successfully constructed，and its genome sequence exhibited high consistency with the sequence of the parental strain. By transfecting target cells，a derivative virus with infectious activity and replication capability was successfully rescued. The derived virus maintained phenotypic characteristics consistent with the parental strain，such as cell tropism and syncytium inducibility.Conclusion:This study successfully constructed an infectious clone of a Chinese HIV-1 CRF01_AE epidemic strain with clear background and distinct phenotype. The genomic sequence and viral phenotypic characteristics of the derived virus are basically consistent with the parental strain，providing strong representation of the original isolate and serving as a powerful tool for research on the correlation between the genetic characteristics，viral phenotype，and pathogenicity of HIV-1 CRF01_AE strains.
