摘要
目的应用假病毒感染实验快速筛选可用于HIV-1中和抗体测定的功能性膜蛋白基因。方法从HIV感染样品中扩増全长膜蛋白基因gp160,通过系统进化树分析确定其基因亚型,将HIV-1gp160阳性克隆与pSG3△Env质粒共转染293T细胞制备假病毒并对其感染能力进行分析,筛选功能性膜蛋白基因。结果克隆和鉴定了我国HIV-1流行毒株的9个功能性膜蛋白基因,系统进化树分析表明其中包括4个CRF07_BC、2个CRF01_AE和3个B亚型毒株,这些gp160膜蛋白基因所制备的假病毒具有良好的感染性。结论不同亚型HIV-1功能性膜蛋白gp160克隆为选择HIV-1中和抗体测定毒株提供了有价值的实验材料。
Objective To rapid screen and characterize functional HIV-1 envelope genes for HIV-1 neutralization test by pseudovirus infection.Methods HIV-1 gp160 genes were amplified from the blood sampls of the infected subjects.HIV-1 clade was determined by sequencing and phylogenetic analysis.HIV-1 pseudoviruses were prepared by transfection of func tional env gp160 clone with plasmid pSG3Env,and the pseudovirus infectivity was also characterized.Results Nine HIV-1 gp160 genes were successfully cloned from HIV-1 infected patients using the HIV-1 pseudovirus infection assay.Sequencing and phylogenetic analysis indicated there were 4 CRF_07BC,2 CRF_01AE and 3 B viruses.Functional analysis demonstrated the pseudoviruses derived from these gp160 clones were infectious.Conclusion The present study has established an assay for rapid screening and characterization of functional gp160 clones of different HIV-1 subtypes,which will be useful in mea surement of HIV-1 neutralizing antibody response.
出处
《中国热带医学》
CAS
2013年第6期659-661,共3页
China Tropical Medicine
基金
"十二五"重大传染病防治研究项目(No.2012ZX10001-008)
国家自然科学基金课题(No.81261120379)
