气肿疽梭菌细胞毒素CctA基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of Cytotoxic CctA gene in Clostridium chauvoei

作　　者：云巾宴 任春宇 车达 金清洙 段雪岩[1] 司唯[1] 金鑫[1] 

机构地区：[1]延边大学农学院动物医学系,吉林延吉133002 [2]延边州畜牧总站,吉林延吉133000 [3]延边州动物疫病预防控制中心,吉林延吉133000 [4]延吉市动物卫生监督所,吉林延吉133000

出　　处：《吉林农业科学》2015年第1期82-85,共4页Journal of　Jilin Agricultural Sciences

基　　金：吉林省教育厅"十二五"科学技术研究项目(吉教科合字[2014]第6号)

摘　　要：根据GenBank中气肿疽梭菌细胞毒素CctA基因的参考序列,采用PCR方法扩增细胞毒素CctA基因。在将该基因进行克隆和测序后利用生物信息学方法,对细胞毒素CctA蛋白的理化性质、结构功能域、蛋白质二级结构和三级结构及抗原表位等重要参数进行了预测和分析。结果表明:与参考序列(JQ692583)相比,存在3处突变,核苷酸序列的同源性和氨基酸序列的同源性均为99.4%;CctA蛋白的理论等电点为8.00,不存在跨膜区和信号肽序列,二级结构以β-折叠和无规则卷曲为主,主要有4个抗原表位区域。本研究为CctA蛋白功能的深入研究提供了参考,并为气肿疽梭菌单克隆抗体的制备及合成肽疫苗的研发奠定了基础。Cytotoxic CctA gene was amplified by PCR according to the GenBank Clostridium chauvoei cytotoxic CctA gene reference sequence. After cloning and sequencing, the cytotoxic CctA protein physical and chemical prop-erties, the structure and function domain, protein secondary structure and tertiary structure and antigen epitope andother important parameters were predicted and analyzed by bioinformatics methods. The result showed that com-pared to the reference sequence（JQ692583） there were three point mutations, the sequence homologies of nucleo-tides and amino acids were both 99.4%. The theoretical isoelectric point of protein was 8.00. Cytotoxic CctA proteindidn＇t exist in the transmembrane region and signal peptide sequence. The secondary structures were mainly in be-ta-folding and random curl, and there were 4 main antigenic epitope regions. This study provided a reference for fur-ther research on cytotoxic CctA protein function, and laid the groundwork for Clostridium chauvoei monoclonal anti-bodies, synthetic peptide vaccine research and development.

关 键 词：气肿疽梭菌 细胞毒素CctA基因 克隆 序列分析 

分 类 号：S852.44[农业科学—基础兽医学]