猪Chemerin和受体基因的克隆与组织分布  

Molecular Cloning and Tissue Distribution of Porcine Chemerin and Its Receptor

作　　者：王静[1] 李宏基[1] 韩立强[1] 王伟杰[1] 武宁晓[1] 张志强[1] 台玉磊[1] 臧猛[1] 杨国宇[1] 

机构地区：[1]河南农业大学动物生理生化实验室

出　　处：《江西农业大学学报》2009年第3期517-521,共5页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金(30671531)

摘　　要：基于电子延伸序列,本试验克隆并分析了猪Chemerin和受体(ChemerinR)基因;各种组织提取总RNA,利用设计的引物进行RT-PCR,PCR产物与pMD19-T连接后转化E.coliJM109,检测阳性克隆并测序;结果表明:猪Chemerin cDNA克隆片段为558 bp,开放阅读框架为492 bp,编码有163个氨基酸。ChemerinR cDNA克隆片段为1 470 bp,开放阅读框架为1 092 bp,编码363个氨基酸。同源分析结果表明,猪Chemerin的核酸序列与牛、人和大鼠的同源性分别为86.7%、79.1%和73%,氨基酸序列的同源性分别为83.1%、83.4%和67.7%,ChemerinR的核酸序列与人、大鼠和小鼠的同源性分别为80.4%、77.6%和72.6%,氨基酸序列的同源性分别为88.2%、80.7%和79.3%。组织分布结果显示:猪Chemerin在多种组织均有分布,ChemerinR仅在膀胱、大脑和肌肉有分布。The coding sequence of porcine Chemerin and its receptor gene were amplified based on the in silico sequence information in the study. Total RNA was extracted from all kinds of organizations and mRNA sequence of gene was amplified by RT - PCR using designed primers. The PCR products were ligated into the pMD - 19T vector, and then transformed into competent cells of E. coli JM109. Porcine Chemerin gene cDNA sequence ,was 558 bp in length, with an open reading frame of 492 bp encoding 163 amino acids. ChemerinR gene cDNA sequence, was 1 470 bp in length, with an open reading frame of 1 092 bp encoding a 363 amino acids. Homology analysis showed that porcine Chemerin and nucleic acid sequences, bovine, human and rat had the homology of 86.7%, 79.1% and 73%, amino acid sequence had the homology of 83.1% ,73.4% and 67.7%. Homology analysis showed that porcine ChemerinR and nucleic acid sequences, human,rat and mouse had the homology of 80.4 % ,77.6% and 72.6% ,amino acid sequence had the homology of 88.2%, 80.7% and 79.3%. Semi - quantitative RT - PCR analysis showed that Chemerin mRNA ubiquitously expressed in various tissues tested, but ChemerinR preferentially expressed in the bladder , brain and muscle.

关 键 词：猪 CHEMERIN 受体 电子克隆 序列分析 组织分布 

分 类 号：S828.212[农业科学—畜牧学]