草鱼(Ctenopharyngodon idellus)NCCRP-1基因的克隆和原核表达  

cDNA Cloning and Prokaryotic Expression of NCCRP-1 from Grass Carp(Ctenopharyngodon idellus)

作　　者：蔡佳[1,2,3] 代礼平[1,2,3] 王蓓[1,2,3] 汤菊芬[1,2,3] 黄郁葱[1,2,3] 鲁义善[1,2,3] 吴灶和[2,3,4] 简纪常[1,2,3] 

机构地区：[1]广东海洋大学水产学院,湛江524088 [2]广东省水产经济动物病原生物学及流行病学重点实验室,湛江524088 [3]广东省水产经济动物病害控制重点实验室,湛江524088 [4]仲恺农业工程学院,广州510225

出　　处：《生物技术通报》2013年第11期105-111,共7页Biotechnology Bulletin

基　　金：国家"973"计划项目(2009CB118704)

摘　　要：应用RACE PCR克隆了草鱼NCCRP-1基因。结果表明,NCCRP-1cDNA全长900 bp,开放阅读框为714 bp,编码237个氨基酸残基,预测分子量为27.3 kD,理论等电点为5.63。草鱼NCCRP-1具有NCCRP-1蛋白家族保守的功能区域,与鲤鱼该基因(Cyprinus carpioL.)的相似性为88%。将此基因定向插入原核表达载体pET-32a,构建重组质粒pET-NCCRP后转化BL2(1DE3)进行诱导表达。SDS-PAGE分析显示在47.8 kD处出现特异性蛋白条带,Western blot检测表明草鱼NCCRP-1融合蛋白成功表达。In the present study, grass carp （ Ctenopharyngodon idellus ） NCCRP-1 gene was cloned using RACE method. The full length cDNA of NCCRP-1 was 900 bp containing a 714 bp open reading frame （ ORF ）, which encoded 237 amino acids with predicted molecular mass of 27.3 kD and a theoretical pI of 5.63. Amino acid alignment indicated that grass carp NCCRP-1 possessed consensus functional domians of NCCRP-1 family and shared 88% similarity with common carp NCCRP-1, The prokaryotic expression vector pET-NCCRP was constructed and then transformed into BL21 （ DE3 ） . SDS-PAGE and Western blotting analysis indicated that the recombinant protein of grass carp NCCRP-1 was successfully expressed and molecular weight of expressed fusion protein was 47.8 kD.

关 键 词：草鱼 NCCRP-1 克隆 原核表达 

分 类 号：S917.4[农业科学—水产科学]