HIV-1B/C重组型外膜蛋白env基因序列分析及表型预测  

Genetic analysis and phenotype prediction of the complete env genes of HIV-1B/C

作　　者：汪孟冉 陈丹瑛[1] 何小周[2] 叶景荣[1] 余双庆[2] 徐柯[2] 曾毅[2] 冯霞[2] 

机构地区：[1]北京工业大学生命科学与生物医学工程学院病毒与药理室,100022 [2]中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室,100052

出　　处：《中华实验和临床病毒学杂志》2014年第3期203-205,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的 克隆并分析HIV-1B/C重组型外膜蛋白env基因序列,根据其氨基酸序列进行表型预测,为疫苗的抗原设计奠定基础.方法 采集北京地区HIV-1 B/C重组型的抗凝全血标本,分离血浆和提取基因组DNA,采用巢式PCR方法扩增rev-env基因,对扩增产物进行序列测定.根据核苷酸序列推导出相应的氨基酸序列,并对重要的Env功能结构域进行深入的分析和比较.结果 从12例B/C重组型HIV-1感染者中成功克隆到7个rev-env基因,序列分析发现其中6个有完整的开放读码框（ORF）,全部为CRF_07B/C重组型.6个Env蛋白氨基酸N糖基化位点和数目没有显著变化;CD4受体结合位点高度保守;根据V3环氨基酸序列及静电荷数目预测全部使用CCR5辅助受体;GP120/GP41剪切位点高度保守,预测所有GP160前体都能有效剪切;对几个已知中和抗体的中和位点分析推测全部的6个序列都对2G12、2F5中和不敏感;对4E10、PG9及PG16中和敏感.结论 有必要进一步阐明env基因型与相关功能的关系,这将为疫苗和药物研究提供依据.Objective To clone and analyze HIV-1B/C env gene sequences,predict their phenotypes according to the deduced amino acid sequences and provide basis for HIV vaccine development.Methods Anticoagulation whole blood samples were collected from HIV-1 B/C infected individuals in Beijing,the env gene fragments were amplified from genome DNA template extracted from plasma using nested PCR methods and the amplified full-length genes were sequenced.The amino acid sequences of these env genes were deduced according to their nucleotide sequences and extensive analysis and comparison of important structural motifs were performed.Results Seven full-length rev-env genes were obtained from 12 HIV-1 B/C infected individuals,sequence analysis found that 6 of them had intact open reading frame （ORF）.All the 6 sequences were conformed to be subtype CRF_07B/C.There was no drastic alteration in the number and position of potential N-linked glycosylation sites among these 6 sequences.And the residues involved in forming the CD4 binding site were highly conserved.Genotypic prediction of coreceptor usage based on V3 sequence and net charge suggested that all samples use CCR5 coreceptor.The cleavage site of Gp120/Gp41 was highly conserved,so Gp160 precursor of all isolates would be efficiently cleaved into the Gp120 and Gp41 subunits.Analysis of several known neutralizing antibody binding sites indicated that the 6 isolates will be insensitive to neutralization by 2G12 and 2F5,but sensitive to neutralization by 4E10,PG9 and PG16.Conclusion Further study to elucidate the correlation of the env genotype to functionally relevant motifs is necessary and that will aid vaccine and novel drug design.

关 键 词：HIV-1 序列测定 基因 env 

分 类 号：R373[医药卫生—病原生物学]