人单核细胞趋化蛋白受体5的基因克隆及该基因超家族成员氨基酸结构

目的：获取有重要生物功能的人单核细胞趋化蛋白受体５（ｈｕｍａｎ ｍｏｎｏｃｙｔｅ ｃｈｅｍｏｔａｃｔｉｃ ｒｅｃｅｐｔｏｒ ５，ｈｕＣＣＲ５）。方法：从人ＰＢＭＣ中提取总ＲＮＡ和ｐｏｌｙ（Ａ）＾＋ＲＮＡ，而后经反转录合成ｃＤＮＡ第一链，再以ＰＣＲ扩增出ｈｕＣＣＲ５ ｃＤＮＡ并插入融合表达载体ｐｃＤＮＡ３．０的ＢａｍＨⅠ和ＨｉｎｄⅢ位点，转化大肠杆菌ＴＧ－１，挑取克隆，酶切鉴定，序列分析。结果：克隆出长度为１０５６ｂｐ，编码３５２个氨基酸的ｈｕＣＣＲ５ ｃＤＮＡ。并分析证明与该基因超家族氨基酸有８０％同源性，与国外发表资料比较有３个碱基突变。结论：克隆出人单核细胞趋化蛋白５受体，为今后基础研究提供了较有用的材料。

Clone of human monocyte chemotactic receptor 5 and amino acid structure analysis of superfamily

Objective:To acquire a human monocyte chemotactic receptor 5(huCCR5) which have important biological function.Methods:Total RNA and poly (A)+ RNA were obtained from human PBMC,and transciptionally synthesized the first strand of cDNA,then amplified huCCR5 cDNA by PCR and inserted it into BamH Ⅰ and Hind Ⅲ site of fusion expression vector,pcDNA 3.0.Sequencely,the vector was transformed into TG-1.Positive clone were selected by enzyme digestion and identification,then the sequence was analyzed.Results:A huCCR5 cDNA of 1 056 bp encoding 352 amino acids was cloned.It had 80% homology with its superfamily and had 3 base mutation compared with reference. Conclusion:A huCCR5 cDNA is cloned and materials are provided for research in the future.