论文部分内容阅读
目的:研究中国HIV-1 E亚型代表株的主要结构基因及其功能。方法:选择3份根据HIV-1 的外膜基因(C2-V3)的序列分析判定为E亚型的阳性血样,采用克隆和系统树分析,通过套式PCR得到全长的gp120 基因片段,并插入到pFastBacl载体中,以双脱氧末端终止法测定全部的DNA 序列。结果:来自广东和广西的样品的gp120 基因归于E亚型,E亚型中国株内的遗传距离为2.56% ～5.87% ,与16 株国际标准株比较,遗传距离为3.60% ～27.98% 。所有克隆到的gp120 基因都具有完整的阅读框架,无大的缺失和插入。结论:HIV-1 E亚型进入中国的时间不长;克隆到的E亚型代表株的gp120 基因具有完整的结构和功能,适合用来构建E亚型的亚单位疫苗表达载体
Objective: To study the major structural genes and their functions of HIV-1 E subtype in China. METHODS: Three positive blood samples of type E subtype were selected from the sequence analysis of the outer membrane gene of HIV-1 (C2-V3). Cloning and phylogenetic tree analysis were used to obtain the full-length gp120 gene fragment by nested PCR Inserted into the pFastBacl vector, and the entire DNA sequence was determined by the dideoxy terminator method. Results: The gp120 gene of Guangdong and Guangxi samples belonged to E subtype. The genetic distance in Chinese isolates of E subtype was 2.56% -5.87%. Compared with 16 isolates, the genetic distance was 3.60% -27.98%. All cloned gp120 genes have a complete reading frame, no major deletions and insertions. CONCLUSION: HIV-1 E subtype enters China for a short period of time. The gp120 gene of the cloned E subtype representative strain has complete structure and function and is suitable for constructing subunit vaccine expression vector of E subtype