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作 者:王斌[1] 钱冬萌[1] 傅继华[2] 鲁晓晴[1] 闫志勇[1] 方荣[1] 吴虹[1]
机构地区:[1]青岛大学医学院微生物学教研室,青岛266021 [2]山东省卫生防疫站
出 处:《青岛大学医学院学报》2003年第1期1-3,共3页Acta Academiae Medicinae Qingdao Universitatis
基 金:中华人民共和国卫生部科研基金资助项目( 9613 5 3 ) ;山东省优秀中青年科学家奖励基金资助项目 ( 973 14 43 2 )
摘 要:①目的 对 1997~ 2 0 0 0年来自华东地区 5 2例HIV 1分离株env基因C2~V3区序列及其V3环顶端主要中和抗原决定簇 (PND)氨基酸序列的变化规律进行分析。②方法 以来自同时期上述地区的HIV 1感染者PBMC基因组为模板 ,采用套式PCR扩增HIV 1env基因C2~V3区片段 ,产物经克隆、测序。DNA测序结果经DNAStar软件分析HIV 1分离株的基因分型 ,比较HIV 1膜蛋白V3环顶端PND相关八肽氨基酸及其编码核苷酸序列的变化特征及出现频率并测其共享序列。③结果 1997~ 2 0 0 0年华东地区HIV 1分离株的基因分型有 7个 ,分别属于HIV 1M亚群的A~G亚型。分离株间的基因离散率为 1.2 %~ 5 .8%。同时发现了env基因双V3区变异的HIV 1自然突变株 (GenbankAF2 2 0 2 4 5 )。分析 5 2株PND相关八肽基因序列 ,其中以第 2、3、7位氨基酸及其编码核苷酸的变化最为显著 ,分别为 11.5 %、9.6 %、9.6 % ,且 3组共享序列相关的八肽亲水性有明显差异。④结论 1997~ 2 0 0 0年华东地区HIV 1流行呈现多个型别共同、混合流行的特征 ,膜蛋白PND的八肽基因序列呈现多样性变化 ,新的HIV 1膜蛋白双V3区变异株的发现可能与病毒重组。Objective\ To analyze the variation of amino acid sequences of principal neutralizing domains(PND) on the V3 loop of env gene C2-V3 fragments from 52 HIV 1 isolates in east China during 1997-2000. \ Methods\ Nested primers PCR was performed to amplify the HIV 1 env gene C2-V3 fragments with the PBMC genome as the templates. The PBMC samples were from 52 HIV 1 sufferers in east China during 1997-2000. The amplified PCR products were cloned into T A vector and by DNA sequencing. The gene type of HIV 1 isolates was confirmed by analysis of DNA sequencing. The consensus sequences and comparison of PND related octapeptide on V3 loop of membrane protein of HIV 1 isolates were performed by DNA Star software. Results\ There were seven gene types among the 52 HIV 1 isolates from east China during 1997-2000. They belonged to A-G subtype of HIV 1M sub group. The divergence of gene among the isolates was 1.2%-5.8%. A natural HIV 1 variant strain (Genbank AF220245) was also discovered, showing double V3 repeated insertion. The prominent variation could be found on the second amino acid (11.5%), the third (9.6%),and the seventh (9.6%). Meanwhile, obvious difference of hydrophilicity could be found among the three groups of consensus octapeptides.\ Conclusion\ The characteristic of epidemic HIV 1 strains in east China during 1997 to 2000 was multi subtypes and mixed prevalence. The octapeptide of PND on membrane proteins showed great changes. The natural mutant HIV 1 strain with double V3 repeated insertion in env gene may relate to natural recombinant, immune escape and mechanism of viral pathology.
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