中国人HIV辅助受体CXCR4基因的克隆及序列测定

Cloning and Sequence Analysis of Chinese CXCR4(a Coreceptor for HIV Infection) cDNA

为了获得ＨＩＶ辅助受体ＣＸＣＲ４基因，进一步探索艾滋病（ＡＩＤＳ）的诊断、防治方法，采用逆转录巢式聚合酶链式反应（ＲＴ－ｎＰＣＲ）从中国人外周血淋巴细胞中克隆了ＣＸＣＲ４ ｃＤＮＡ基因，得到了预期的１ １００ｂｐ左右的片段。将ＰＣＲ产物与Ｔ载体连接，并进行了酶切反应鉴定，获得与Ｔ载体正向连接的克隆。测序结果表明：克隆的片段含有一个开放性阅读框架，编码３５２个氨基酸残基。搜索ＧｅｎＢａｎｋ，目标片段与Ｍ９９２９３、ＸＭ０５１２２３、ＸＭ０５１２２４、 ＸＭ０５１２２５有很高的同源性，比较结果显示：克隆的片段包括完整的编码序列，含两个碱基的差异。

In order to acquire the CXCR4 gene and develop th e method to diagnose and treat AIDS patient, the cDNA of CXCR4 was cloned by RT－nPCR from peripheral blood mononuclear cells of a Chinese. An expected target fragment was identified by restriction enzyme digestion and DNA sequence data. The sequen ce data showed that CXCR4 gene is 1 059 nt encoding 352 amino acids. Compared wi t h the sequence from GeneBank, there were only two deferent sites. Searched in Ge nBank, it was found that the segment had high homologenicity with M99293、XM051 223、XM051224、XM051225. The comparison result showed that the cloned segment included intact coding sequence and two basepair deference.