早期HIV-1感染env基因的动态变异研究  

Dynamic Variation of HIV-1 CRF07_ BC env gene in early HIV-1 infection

作　　者：刘胜牙[1] 邢辉[1] 张远志[3] 阮玉华[1] 全宇[1] 朱家鸿[2] 邵一鸣 

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心传染病预防控制国家重点实验室艾滋病组,北京100050 [2]武汉生物制品研究所 [3]新疆疾病预防控制中心

出　　处：《中华实验和临床病毒学杂志》2008年第4期290-292,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家十五科技攻关项目（2004BA719A01）;国家重点基础研究发展计划（973计划）2005CB522903

摘　　要：目的追踪观察HIV-1新发感染者体内CRF07-BC重组毒株膜蛋白基因的变异性。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全长及C2-C5区段基因。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，运用PCR方法进行鉴定，最后对所获得的目的克隆测序。结果从两个感染者血浆样品中获得感染后半年到2年半间多个时间点的gp120基因克隆共135个及gp120基因C2-C5区段克隆15个，分析显示这些克隆均为HIV-1cRFlD7_BC亚型。随感染时间的延长，HIV-1env基因的离散率与多样性均有增加的趋势。env基因同义突变与非同义突变比较结果显示，C1、C3与V4区段非同义突变比率比gp120基因的其他区域都高。基因多态性分析的结果显示，同-时间点内不同毒株基因在不同区段的变异是不同的，基因多样性介于0与0．066±0．028之间。结论随着患者感染时间的推移，HIV-1膜蛋白基因的变异逐渐增大。C1、C3和V4区的高突变率提示，该基因区可能是HIV-1病毒生存与宿主免疫压力相互作用的主要位点。Objective To understand the evolution of HIV-1 CRF07_ BC envelope, we performed a longitudinal study on two patients during their early HIV-1 infection. Methods RNA was extracted from the plasma of the individuals and the C2-C5 fragments of the gpl20 gene of HIV-1 were amplified by RT-PCR. Purified DNA segments were inserted into T easy vector and transformed into E. coli Topl0 competent cells. Positive clones were identified by blue-white screening, confirmed by PCR and sequenced by ABI 3700. Results The samples were collected from the patients every 6 months from seroconversion time. The genetic diversity and divergence in env gene showed consistent increases over time. Our sequence analysis also revealed obvious non-synonymous change in env C1, C3 and V4 regions among these samples. Conclusion The results support the concept that the consistent pattern of viral evolution existed during eady phase of HIV-1 infection. C1, C3 and V4 region of env gene may be mainly immunological target during AIDS progression.

关 键 词：HIV-1 准种 多样性 离散率 变异(遗传学) 

分 类 号：R512.91[医药卫生—内科学]