犬弓首蛔虫enol-1基因的克隆及序列分析  

Cloning and Sequence Analysis of Toxocara canis enol-1 Gene

作　　者：卜卜才加 张念章[2] 张芙恺 朱兴全[2] 赵权[1] BUBU Caijia;ZHANG Nianzhang;ZHANG Fukai;ZHU Xingquan;ZHAO Quan(College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;State Key Laboratory of Veterinary Etiological Biology,Key Laboratory of Veterinary Parasitology of Gansu Province,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences ,Lanzhou 730046,China)

机构地区：[1]吉林农业大学动物科学技术学院,长春130118 [2]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室,兰州730046

出　　处：《吉林农业大学学报》2019年第1期97-101,共5页Journal of Jilin Agricultural University

基　　金：中国农业科学院基本科研业务费专项(Y2016JC05;1610312017004);中国农业科学院科技创新工程项目(CAAS-ASTIP-2014-LVRI-03)

摘　　要：对犬弓首蛔虫烯醇化酶(enolase)基因(enol-1)进行克隆及序列分析,预测其作为抗犬弓首蛔虫疫苗候选抗原的潜力。从犬弓首蛔虫的cDNA中扩增出enol-1基因,连接至pMD18-T载体,并筛选阳性克隆后,测序鉴定。将测序结果正确的核苷酸序列翻译成氨基酸序列,运用生物信息学软件分析enol-1蛋白的结构,预测抗原表位及功能位点,并运用Maximum likelihood(ML)法构建了系统发育树。结果表明:enol-1基因长1 311 bp,可编码437个氨基酸。Enol-1蛋白中包含18个α-螺旋区,7个β-折叠区,10个亲水区域和16个柔性区域,并预测含有9个线性B细胞抗原表位,enol-1蛋白可能具有潜在的免疫原性。并对犬弓首蛔虫enol-1基因进行了克隆及序列分析,为评价enol-1蛋白作为犬弓首蛔虫亚单位疫苗及表位疫苗奠定了基础。In order to predict the possibility of Toxocara canis enolase gene(enol-1) as anti-T. canis vaccine candidate, enol-1 gene was amplified from cDNA of T. canis. The fragment was cloned into pMD18-T vector, and the positive clones were screened and sequenced. The correct nucleotide sequence was translated into amino acids and analyzed by bioinformatics analysis. The epitopes and functional sites were predicted, and the phylogenetic tree was established. The results revealed that enol-1 gene was 1 311 bp in length, encoding 437 amino acids. Enol-1 protein was predicted to contain 18 α-helix, 7 β-fold, 10 hydrophilic regions, 16 flexible regions and 9 linear B cell epitopes, indicating that enol-1 can be a vaccine candidate antigen against T. canis. In this study, the enol-1 gene of Canine aphid has been cloned and sequenced for the first time, which will lay a foundation for the evaluation of enol-1 protein as a subunit vaccine and epitope vaccine.

关 键 词：犬弓首蛔虫 enol-1基因 克隆 生物信息学分析 

分 类 号：S852.7[农业科学—基础兽医学]