草鱼穿孔素C端溶细胞活性分析  

Cytolytic Activity Analysis of Grass Carp Perforin C-terminal Peptide

作　　者：夏玉洁[1] 吴初新[1] 胡成钰[1] 

机构地区：[1]南昌大学生命科学与食品工程学院,南昌330031

出　　处：《中国生物工程杂志》2010年第1期47-50,共4页China Biotechnology

基　　金：江西省重点科技攻关项目(20061B0260301)资助项目

摘　　要：穿孔素的溶细胞作用是机体杀伤病毒和其他微生物感染细胞以及肿瘤细胞的一种效应机制。穿孔素在鱼类非特异性免疫中起重要作用。为了解鱼类穿孔素的功能,根据穿孔素基因序列特征,在已构建的草鱼肝肾cDNA文库中克隆了草鱼穿孔素基因C端包含1个完整蛋白激酶C保守结构域(C2)的cDNA。将该cDNA与表达载体pET32a连接并转化表达菌DE3,诱导表达。His-Bind亲和柱纯化获得了草鱼穿孔素C端表达多肽(PFP-C)。将PFP-C与兔红细胞共育,结果表明:PFP-C具有溶血功能,且在pH7.5时活性最大,其溶血活性对Ca2+有明显的依赖性。The cytolytic effect of perforin is a mechanism of anti-virus,killing microbial-infected cells and tumor cells.Perforin is a very important non-specific immune factors in fish.In order to understand the function of perforin,the cDNA of grass carp perforin C-terminal peptide was amplified from grass carp liver and kidney cDNA library.It contains a protein kinase C conserved region 2（C2）.The cDNA was connected with pET32a,and transformed to expression bacteria DE3.PFP-C was expressed by a prokaryotic expression system and then purified by affinity chromatography.It showed a significant haemolytic activity when tested with rabbit red cells,the optimal pH for haemolytic activity was 7.5,and its haemolytic function dependents on Ca^2＋ apparently.

关 键 词：穿孔素 C2 溶细胞 草鱼 

分 类 号：Q785[生物学—分子生物学]