| 梁浩,邢辉,魏民,陈钊,关琪,黄海龙,全宇,陈健平,洪坤学,施侣元,邵一鸣.中国艾滋病病毒1型AE循环重组型毒株env基因的变异和进化分析[J].Chinese journal of Epidemiology,2003,24(11):966-970 |
| 中国艾滋病病毒1型AE循环重组型毒株env基因的变异和进化分析 |
| Study on the evolutionary pressure on the env gene of the human immunodeficiency virus type 1 CRF01-AE strains circulating in China |
| Received:January 28, 2003 |
| DOI： |
| KeyWord: 人类免疫缺陷病毒 基因 变异 进化,分子 |
| English Key Word: Human immunodeficiency virus Gene Variation Evolution, molecular |
| FundProject:2000年度国家科技部基础性工作专项资金资助项目(120);国家重点“973”基础研究资助项目(G1999054107) |
| Author Name | Affiliation | | LIANG Hao | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | XING Hui | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | WEI Min | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | CHEN Zhao | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | GUAN Qi | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | HUANG Hai-long | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | QUAN Yu | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | CHEN Jian-ping | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | HONG Kun-xue | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China | | SHI Lv-yuan | 华中科技大学同济医学院公共卫生学院 | | SHAO Yi-ming. | National Center for Sexually Transmitted Disease and Acquired Immune Deficiency Syndrome Prevention and Control, Chinese Center for Disease Prevention and Control, Beijing 100050, China |
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| Abstract: |
| 目的研究中国艾滋病病毒1型(HIV-1)AE循环重组型(CRF01-AE)毒株env不同基因区序列变异的特点及其与进化压力的关系.方法应用巢式聚合酶链反应(nested-PCR)对从全国部分省收集来的HIV-1感染者血液样本中的HIV-1 外膜蛋白(env)基因进行扩增和亚型鉴定后,选择34份CRF01-AE重组型HIV-1毒株env基因V3～V4区及其邻近区域的序列进行系统进化树和氨基酸变异分析,并计算和分析氨基酸同义替换(Ks)值和非同义替换(Ka)值及Ks/Ka比值.结果基因系统树显示中国的34份样本CRF01-AE重组型毒株与我国代表株AE.97CNGX2F和泰国代表株AE.CM240、AE.93TH253聚集在一起.氨基酸替换主要发生在C3和V4区,而V3区和V3上游区氨基酸序列相对保守,糖基化位点也比较保守.V3环顶端四肽以GPGQ为主(87.50%).大部分毒株的第306和320位点上没有出现带正电荷的氨基酸.整个V3～V4区的Ks值显著高于Ka值(P＜0.001),且Ks/Ka比值显著高于1(P＜0.001),只有V4区Ks/Ka比值显著低于1(P＜0.01).结论目前多数流行于中国的CRF01-AE重组型HIV-1毒株具有较高的同源性,在进化上关系密切.氨基酸替换主要发生在C3和V4区,而不是V3区.由于大部分毒株的第306和320位点上没有出现带正电荷的氨基酸,推测这些毒株为不引起细胞融合的NSI型,其V3～V4区序列的变异主要受到功能性的限制而与正向选择压力无密切关系,但其中V4区的变异与正向选择压力有关. |
| English Abstract: |
| Objective To identify variations in the envgene of human immunode ficiency virus type 1(HIV-1) subtype CRF01-AE strains circulating in China and to elucidate the potential relationship between genetic variation and evolutionary pressure.Methods Fragments of the HIV-1 env gene were amplified by nested-polymerase chain reaction(n-PCR)from the whole blood of HIV-1 infected individuals f rom four provinces in SoutheasTchina (Guangdong, Hunan, Jiangsu and Jiangxi).The PCR products were then directly sequenced by ABI 377 DNA sequencers.The sequences covering the env V3-V4 region of 34 HIV-1 subtype CRF01-AE strains were selected to analyse phylogenetic trees and amino acid mutations.The accumulation of synonymous (Ks)and antonymous (Ka)substitutions as well as Ks Ka ratios were calculated using DIVERGE.Results Phylogenetic trees showed thaTthe 34 HIV-1 subtype CRF01-AE strains from China clustered with the Chinese AE reference strain (AE.97CNGX2F), as well as with the reference strains from Thailand (AE.CM240 and AE.93TH253).The amino acid sequences of the env V4 and C3 regions in the samples were highly variable, compared with those of V3 and V3-downstream regions.The V3loop central motif in the majority (87.5%)of the strains was GPGQ.The majority of strains did noTcontain positiv ely charged amino acids aTpositions 306 and 320 in V3loop.The N-linked g lyco sylation sites in the V3-V4 region and flanking regions in these strains were relatively conserved.Analysis of the entire region showed thaTthe mean Ks values were significantly higher than that of the Ka values (P <0.001), with the Ks Kasig nificantly higher than 1.0 (P <0.001).In contrast, the Ks Ka ratio in the V4 region was significantlylower than 1.0 (P <0.01).Conclusions Our study indica ted thaTthe majority of HIV-1 subtype CRF01-AE strains circulating in China were highly homogeneous.The amino acid sequences of the V4 and C3 regions were significantly more variable than those of the V3loop.Our analysis also suggested thaTthe phenotype of nearly all strains was likely to be non-syncy tium inducing (NSI).Finally, the variation found in the V3-V4 sequence was significantly influenced by functional constraints as opposed to positive selective pressure, while the variability of thelone V4 region was strongly related to positive selective pressure. |
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