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目的 了解我国HIV感染者p2 4蛋白编码区的基因变异情况。 方法 收集我国河南及上海地区 2 8份已经证实的HIV - 1感染患者的血浆标本并抽提出RNA ;采用逆转录和Nested PCR的方法扩增所需的目的基因 ,使用DNA测序仪直接测序 ;应用CLUSTALW、PHYLIP等软件包对序列进行比对及进化树分析。结果 2 8份gag样本中 2 5份为B亚型 ,3份为A亚型。与共享序列相比 ,2 5例B亚型的 p2 4编码区中发生核苷酸改变的位点比例为 0 .4 %～ 4 .8% ,平均为 1.4 % ,其中A→G占 2 0 .5 % ,G→A占 17.3% ;3例A亚型发生核苷酸改变的位点比例平均为 0 .2 4 %。在所有变异位点中均未发现G→A的高度突变。B亚型和A亚型内的基因离散率平均为 2 .9%和 0 .5 8% ,A亚型与B亚型之间的基因离散率平均为 11.1%。 2 5例B亚型根据基因序列所推测的p2 4蛋白发生氨基酸改变的比例为 0 .4 %～ 5 .2 % ,平均为 2 .2 %。进化树分析表明在我国河南地区HIV感染B亚型多为与泰国株相近的B’亚型。结论 HIV 1p2 4编码区仍相对保守 ,选择无变异发生的区域有助于免疫治疗及疫苗研制等方面的研究
Objective To understand the genetic variation of p2 4 protein coding region in HIV-infected people in China. Methods Plasma samples from 28 confirmed HIV - 1 infected patients in Henan and Shanghai areas of China were collected and RNA was extracted. The desired gene was amplified by reverse transcription and Nested PCR and sequenced directly by DNA sequencing. CLUSTALW, PHYLIP and other software packages for sequence alignment and evolutionary tree analysis. Results 25 of the 28 gag samples were subtype B and 3 were subtype A. Compared with the shared sequences, the proportion of nucleotide changes in the p2 4 coding region of 25 subtype B subtypes was 0.4% -4.8% with an average of 1.4%, among which A → G accounted for 20 .5%, G → A accounted for 17.3%; 3 cases of A subtypes nucleotide changes in the average ratio of 0.24%. No mutation of G → A was found in all the mutation sites. The gene dispersions in subtype B and subtype A averaged 2.9% and 0.58%, respectively, and the average genetic divergence between subtype A and subtype B was 11.1%. According to the deduced gene sequence, the percentage of amino acid changes of p2 4 protein in 25 cases of subtype B was 0.4% ~ 5.2% with an average of 2.2%. Phylogenetic tree analysis showed that most of the B subtypes of HIV infection in Henan were similar to the B ’subtype of Thai strain. Conclusion The coding region of HIV 1p2 4 is still relatively conservative. The selection of non-mutated regions contributes to the research of immunotherapy and vaccine development