黄喉拟水龟巨噬细胞炎症蛋白-3α基因的序列特征与组织表达  

Analysis of gene sequence characteristics and tissue expression on macrophage inflammatory protein-3α in Mauremys mutica

作　　者：龙建杰[1,2] 朱新平[1,2] 史燕[1] 赵密[2] 刘阳[1,2] 

机构地区：[1]中国水产科学研究院珠江水产研究所/农业部热带亚热带水产种质资源利用与养殖重点实验室,广州510380 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《华中农业大学学报》2013年第5期112-118,共7页Journal of Huazhong Agricultural University

基　　金：国家农业科技成果转化资金项目(2011GB23260021);中国水产科学研究院院级基本科研业务费专项(2012A0403);广州市珠江科技新星专项(2012089)

摘　　要：利用已构建的黄喉拟水龟(Ma)SMART全长cDNA文库,筛选得到巨噬细胞炎症蛋白-3α(MIP-3α)全长cDNA序列,通过设计引物、克隆测序验证,采用相关软件对基因的结构与功能进行预测,并利用实时荧光定量PCR对MaCCL20组织表达特征进行分析。序列结构分析结果表明,MaCCL20 cDNA全长2 318 bp,开放阅读框为261 bp,共编码86个氨基酸,其蛋白为疏水性,不存在跨膜结构。同源性分析表明,黄喉拟水龟CCL20与变色蜥(Anolis carolinensis)亲缘关系最近,与原鸡(Gallus gallus)其次,与哺乳动物的同源性很低;荧光定量PCR结果表明,MaCCL20在肝脏、肾脏、心脏、脾脏中均有表达,其中脾脏中表达量最高。Macrophage inflammatory protein-3α（MIP-3α or CCL20）,a member of the chemokine family,plays an important role in the immune response.In this study,the Mauremys mutica CCL20（Ma CCL20） cDNA sequence was initially obtained by screening the constructed full-length cDNA library and were verified by gene cloning method.Then the structure and function of the MaCCL20 gene were predicted by using software and the gene expression in different tissues was analyzed by using real-time quantitative PCR.The results showed that the full-length cDNA of MaCCL20 gene is 2 318 bp,containing a 261 bp open reading frame and encoding 86 amino acids.The MaCCL20 protein is hydrophobic and has no trans-membrane structure.Homology analysis indicated that the MaCCL20 shared high similarity with Anolis carolinensis,followed by Gallus gallus and shared the lowest similarity with mammalian.Real-time PCR results showed that the MaCCL20 gene was highly expressed in the spleen and was also expressed in other tissues including liver,kidney and heart.The study laid a foundation for further research of the function of MaCCL20.

关 键 词：黄喉拟水龟 巨噬细胞炎症蛋白-3α CDNA文库 序列分析 组织表达 

分 类 号：S947[农业科学—水产养殖]