HIV1膜蛋白PND编码基因自然变异株的发现

目的 对1例来自华东地区HIV1分离株（WWBH7）的前病毒env基因C2-V3区进行序列分析。方法 以HIV1感染者外周血单个核细胞基因组为模板进行套式PCR扩增HIV1env基因C2-V3区片段，将此扩增产物插入T-Vector，酶切鉴定重组质粒，使用AB1737自动DNA序列测定仪测定序列并用DNASIS软件进行分析。结果 DNA序列资料显示该毒株属于HIV1B亚型衍生株，但与HIV1B亚型的标准株如SF2株相比，该HIV1毒株的env基因V3区下游有192bp的重复插入突变，使得该毒株的膜蛋白PND编码基因呈现双V3区的变异该段DNA序列已登录于GenBank(AF220245)。结论 该分离株是1个在膜蛋白PND编码基因有大片段插入突变的HIV1变异株。

Discovery of natural variant strain of HIV 1 in env gene c2-v3 encoded PND of membrane protein

Objective To sequence and analyze the env gene C2 V3 region of proviral genome from a HIV1 isolate(WWBH7) which was obtained from Huadong Area in China.Methods The env gene C2 V3 DNA fragment was amplified by nested primer PCR with genome DNA of peripheral blood mononuclear cells from a confirmed HIV1 infected individual as a template. The amplified DNA fragment was inserted into pGEM T vector. The recombinant plasmid was confirmed by restriction enzyme analysis. The inserted DNA fragment was sequenced by ABI737 autosequencer and analyzed by PROSIS software.Results The HIV1 strain was the derivatives of HIV1 B subtype. But there was mutation of 192 bp fragment repeated insertion at env C2 V3 region of the HIV1 strain compared with standard HIV1 B subtype such as SF2 strain. The mutation brought about a double V3 region in gene encoded PND (principal neutralizing domains). The DNA sequence was registered in GenBank (AF220245).Conclusion This was a natural mutated variant strain of HIV1 whose genome showed a 192 bp repeated insertion at C2 V3 region of env gene encoded PND of membrane protein.