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目的通过研究人类免疫缺陷病毒1型(human immunodeficiency virus1,HIV-1)感染者个体内准种间的差异,探讨HIV的系统进化的发生模式。方法从HIV-1感染者血浆中提取总RNA,通过逆转录多聚酶链反应(RT-PCR)获得HIV-1gp120全长基因,纯化后装入T载体,转化至ToP10大肠埃希菌内增殖,通过蓝白斑筛选获得阳性克隆,对所获得的目的克隆测序并分析。结果获得同一患者的16个克隆的gp120全长基因序列,通过系统进化树分析,克隆序列均为CRF07-BC亚型,但在系统进化树上16个克隆可明显分为A、B两群,其中13个克隆属于A群,2个克隆属于B群,1个克隆(编号XPD7)位于A、B群之间,通过simplot软件的重组分析,发现XPD7克隆为A群和B群的重组株。结论发现了我国广泛流行的HIV-1CRF07-BC毒株准种间的重组现象,准种间的重组作为HIV进化的一种有效手段将导致HIV毒株的快速进化的发生,可能更易逃脱宿主的免疫监控。
OBJECTIVE: To investigate the mode of phylogenetic evolution of HIV by studying the inter-species differences among individuals infected with human immunodeficiency virus 1 (HIV-1). Methods The total RNA was extracted from the plasma of HIV-1 infected persons and the full-length HIV-1 gp120 gene was obtained by reverse transcription polymerase chain reaction (RT-PCR). After purification, the full-length HIV-1 gp120 gene was transformed into ToP10 E.coli and propagated Positive and negative clones were obtained by blue and white screening, and the obtained clones were sequenced and analyzed. Results The full-length gp120 gene sequences of 16 clones of the same patient were obtained. The cloned sequences were all CRF07-BC subtype by phylogenetic tree analysis. However, 16 clones in the phylogenetic tree could be divided into A and B groups, Among them, 13 clones belonged to group A, 2 clones belonged to group B, and 1 clone (number XPD7) was located between group A and group B. Recombinant XPD7 was found to be a recombinant strain of group A and group B by means of recombination analysis of simplot software. Conclusions The recombination between the quasi-species of HIV-1CRF07-BC strain, which is widely prevalent in our country, was found. The quasi-interspecific recombination as an effective means of HIV evolution will lead to the rapid evolution of HIV strains and may escape the host more easily Immune monitoring.