山羊NLRP3基因的克隆及其在DF-1细胞中的表达  

Cloning and expression-in-DF-1 cells of NOD-like receptor protein 3 gene of goat

作　　者：张凯照 叶锦辉[1] 于梦[1] 王晴楠 刘健新[1] 陶攀[1] 亓文宝[1] 宁章勇[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医科学》2015年第4期423-428,共6页Chinese Veterinary Science

基　　金：广东省自然科学基金项目(s2013010013041);广东省高等学校优秀青年教师培养计划项目(Yq2013025)

摘　　要：为了解山羊NLRP3基因及其编码蛋白的生物学信息,采用RT-PCR扩增了海南黑山羊NLRP3基因并进行了序列及其编码蛋白的结构分析,构建了pIRES2-EGFP-NLRP3真核表达载体,并将其转染至DF-1细胞中进行了表达。结果表明,NLRP3基因在物种内的同源性高,种间同源性低,编码的蛋白由1 031个氨基酸组成,存在3个潜在跨膜区,分别为第401~419、435~453和884~904位氨基酸,不含信号肽;辣根过氧化物酶单层细胞免疫化学检测证实,构建的真核表达载体pIRES2-EGFP-NLRP3在DF-1细胞中成功表达。本研究结果为建立NLRP3专性表达细胞系及深入研究NLRP3在山羊抗感染免疫的机制奠定了基础。To comprehensively understand the biological information of NLRP3 gene and its encoding protein,the NLRP3 gene of Hainan Black Goat was amplified by RT-PCR and its sequence and coding protein structure were analyzed.The eukaryotic expression vector pIRES2-EGFP-NLRP3 was constructed and transfected into DF-1cells for expression.In result,the NLRP3 gene shows high homology within species while low similarity among the different species.The NLRP3 consists of 1 031 amino acids and has three potential transmembrane domains which contain 401 to 409,435 to 453and 884 to 904amino acid residues but lack of signal peptide.The horseradish peroxidase monolayer cell immunochemical detection confirmed that eukaryotic expression vector,pIRES2-EGFP-NLRP3,was successfully expressed in DF-1cells.The results provided the basic data for construction of NLRP3 expression cell line and further study of goat antiinfection immune mechanism of NLRP3.

关 键 词：海南黑山羊 NLRP3基因 序列分析 真核表达 

分 类 号：S858.27[农业科学—临床兽医学]