我国中部HIV-1B′亚型分离株基因组结构特征及系统发育分析

Strutural Characterization and PhylogeneticAnalysis of HIV-1 Genome of a Prevalent Subtype B′Strain in Central China

为了分析河南省Humanimmunodeficiencyvirus-1(HIV-1)流行株的基因特征和传染来源,我们从一位HIV-1感染的有偿献血员体内分离HIV-1毒株。提取感染细胞的全基因组DNA,扩增病毒全基因组,Walking法测得全长序列。与HIV-1RL42和HIV-1HXB2株比较,分析基因特征。用Anthewin软件比较分析CNHN24株和RL42株env蛋白的二级结构。用Clustal软件对国内HIV-1分离株和国际参考株的全基因组、env基因和V3环基因分别进行了系统发育分析。结果发现,CNHN24株为HIV-1B′亚型,富含嘌呤,含量达到60.26%。与RL42株及HXB2株比较,pol基因和gag基因变异度较小,env基因及非结构基因变异度较大。与RL42株相比,env蛋白二级结构变异不大,但env保守区C4的氨基酸呈高度变异。系统发育分析显示,CNHN24株与RL42株遗传距离最近,HIV-1Lai株和HXB2株次之,与国内的HIV-1B/C及AE/BC重组毒株的遗传距离较远;从V3序列的比较可以发现,CNHN24株与来自云南省的毒株HIV-1CR206及RL42遗传距离最近。认为HIV-1CNHN24株可能由云南传入。

In order to analyze the gene characterization of Human immunodeficiency virus-1(HIV-1) prevalence in Henan Province, a HIV-1 strain, named CNHN24, was isolated from one HIV-1 infected blood donor. The proviral DNA was extracted from productively infected PBMCs (peripheral blood mononuclear cells) and full-length HIV-1 genomes were amplified. The sequences were determined by Walking sequencing. All of the reading frames were compared with HIV-1 RL42 and HIV-1 HXB2. The secondary structure prediction of the envelope proteins of HIV-1 CNHN24 and RL42 were performed by Anthewin software. Multiple alignments of the whole genome, env and V3 loop gene with domestic HIV-1 strains and standard representatives of HIV-1 were performed by Clustal software package, and phylogenetic tree were constructed. The results showed that the HIV-1 CNHN24 was B′subtype (according to V3 nucleotide sequence). There were more purines in the HIV-1 CNHN24 genome, it was 60.24%. Compared with HIV-1 RL42 and HXB2, the homegeneity of gag and pol reading frames was higher, env and non- structural genes was lower. The envelope proteins secondary structure of CNHN24 and RL4 were homoplastic, but the amino acid variability was higher in conservative C4. Phylogenetic trees showed the closest relationship between CNHN24 and RL42, the next was between HIV-1 Lai and HIV-1 HXB2. HIV-1 V3 sequence phylogenetic tree also indicated the closest relationship of CNHN24 to CR206 and RL42, which were isolated in Yunnan Province. We inferred that the HIV-1 CNHN24 may derive from Yunnan Province.