中国HIV-1B/C重组病毒的gag-pol区基因序列特征分析  

Sequence analysis of gag-pol gene of HIV-1 B/C recombinant viruses in China

作　　者：关琪[1] 魏民[1] 黄海龙[1] 邢辉[1] 洪坤学[1] 马鹏飞[1] 梁浩[1] 司雪峰[1] 黑发欣[1] 张卓然[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与控制中心病毒免疫室,北京100050 [2]大连医科大学微生物学教研室

出　　处：《中华医学杂志》2004年第5期387-391,共5页National Medical Journal of China

基　　金：国家重点基础研究资助项目 (G19990 5 410 7);国家杰出青年资金资助项目 (3 992 5 0 3 0 )

摘　　要：目的　对 6株中国人类免疫缺陷病毒 1型 (HIV 1)B/C重组病毒的完整 gag基因和部分pol基因进行序列分析 ,从基因水平上分析是否存在新模式的B/C重组病毒并与其母本毒株进行比较研究 ,尝试对其不同的生物学表型进行解释。方法　从确诊的HIV感染者的全血样本中 ,提取样本基因组DNA ,经套式聚合酶链反应 (PCR)扩增后 ,将扩增产物进行纯化和测序。然后将所得序列进行系统进化树和氨基酸变异分析。用Simplot软件进行序列重组分析并确定重组断点区域 ;用MEGA软件按断点分段做基因进化树分析以验证该断点的正确性 ;用GCG软件包的Distance程序计算基因距离。并分析所研究的HIV 1B/C重组毒株长 2 5 5 0bp基因区段的分区段的基因离散率及基因重组对其功能可能造成的影响。结果　新疆 5份样本均未发现重组断点的变化 ,而重庆 1份样本的逆转录酶区内的B/C断点发生了 16 0个核苷酸的移动。氨基酸序列分析显示 ,我国流行的B/C重组株与其母本B、C毒株之间发生第 2 86位 (R→K/N)和第 799位 (A→T)的变化。结论　我国现在流行的HIV 1B/C重组病毒仍以CRF0 7 BC和CRF0 8 BC两种模式为主 ,在本研究涉及的基因区内尚未发现新模式重组毒株的流行。初步分析表明我国B/C重组毒株 2 86位和 799位氨基酸的变异可能为B/C重组株在我?Objective To characterize full length gag gene and partial pol gene of Chinese prevalent HIV-1 B/C recombinant strains,to explore the genetic difference between parent strains and B/C recombinant strains, and to investigate the mechanism of different biologic phenotype among them. Methods The peripheral blood samples were collected from 138 HIV-positive persons from 12 regions of China respectively. Samples of total DNA were extracted from the peripheral blood movonuclear cells to undergo nested PCR and sequencing. The 117 HIV-1 CRF07-BC strains and 21 HIV-1 CRF08-BC strains were screened with the resulted that 5 CRF07-BC samples from Xinjiang and 1 CRF08-BC sample from Chongqing were regarde as the most potentail new type recombinant viruses. The sequences thus obtained underwent phylogenetic tree analysis and amino acid variation analysis. Simplot software was used to analyze the sequence recombination and identify the breakpoints of B/C recombinant strains. To confirm the breakpoints, separate phylogenetic analysis according to the breakpoints was performed with MEGA software. The genetic distances of different gene fragments were calculated by DISTANCE program in GCG software package. The gene dispersion in the gene fragment of a length of 2550 bp of the recombinant HIV-1-B/C and the potential influence of gene recombination on its function were further analyzed. Results No change of breakpoint was found in the 5 samples from Xinjiang. But a breakpoint shift of 160 nucleotides occurred in RT region of a sample from Congqing city. Conclusion CRF07-BC and CRF08-BC remain the main prevalent HIV-1 B/C recombinant strains in China. No epidemic of new mosaic recombinant strain is found. The variation of amino acids at the sites 286 and 799 may be the reasons of the transmission dominance of the B/C recombinant strain.

关 键 词：人类免疫缺陷病毒1型 重组病毒 GAG基因 pol基因 生物学表型 

分 类 号：R346[医药卫生—基础医学]