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分析(英文)

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

目的 :获取有重要生物功能的人单核细胞趋化蛋白受体 5 (hum an m onocyte chemotactic receptor5 ,hu CCR5 )。方法 :从人 PBMC中提取总 RNA和 poly(A) + RNA,而后经反转录合成 c DNA第一链 ,再以 PCR扩增出 hu CCR5 c DNA并插入融合表达载体 pc DNA3.0的 Bam H 和 Hind 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 10 5 6 bp,编码 35 2个氨基酸的 hu CCR5 c DNA。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。

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 BamHⅠand HindⅢ 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. [