多头带绦虫硫氧还蛋白过氧化物酶基因的克隆及序列分析  

Cloning and Sequence Analysis of Thioredoxin Peroxidase Gene from Taenia multiceps

作　　者：李永光[1,2] 李文卉[1] 盖文燕[1] 姚菊霞[1] 曲自刚[1] 贾万忠[1] Radu Blaga 付宝权[1] 

机构地区：[1]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,农业部兽医公共卫生重点开放实验室,甘肃省动物寄生虫病重点实验室,兰州730046 [2]甘肃农业大学动物医学院,兰州730070 [3]罗马尼亚农业与兽医大学兽医学院,克鲁日-纳波卡400372

出　　处：《中国寄生虫学与寄生虫病杂志》2011年第1期67-70,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家科技支撑计划项目(No.2007BAD40B04);中央级公益性科研院所基本科研业务费专项(No.BRF080303);中国-罗马尼亚政府间科技合作项目(39-18)~~

摘　　要：从自然感染的羊脑内采集脑多头蚴原头节,提取总RNA。根据亚洲牛带绦虫的TaHc2-D11 mRNA序列设计特异引物,采用RT-PCR技术扩增多头带绦虫硫氧还蛋白过氧化物酶(TmTPx)基因。PCR产物连接到pMD18-T载体构建重组质粒pMD-TmTPx,转化大肠埃希菌DH5α后筛选阳性克隆,经限制性酶切及测序鉴定后进行序列分析。扩增获得大小为614bp的TmTPx基因cDNA,该基因的完整开放阅读框架(ORF,591bp)编码196个氨基酸,编码蛋白的相对分子质量为Mr 21690,等电点为7.61。生物信息学分析结果表明TmTPx具有一个典型的2-Cys Prx保守功能结构域。绦虫已知TPx的分子进化分析发现,多头带绦虫与亚洲牛带绦虫的亲缘关系最近,与猪带绦虫和肥头绦虫的亲缘关系次之,与细粒棘球绦虫和多房棘球绦虫的亲缘关系最远。Protoscoleces of Taenia multiceps were collected from the naturally infected sheep and total RNA was extracted.Specific primers were designed according to TaHc2-D11 mRNA sequence and T.multiceps thioredoxin peroxidase gene（TmTPx） was amplified by RT-PCR.PCR products were ligated into pMD18-T vector and transformed to E.coli DH5α.The recombinant plasmids were identified by restriction digestion and sequencing.A 614 bp cDNA was amplified.The TmTPx open reading frame（591 bp） encoded a 196-amino acid protein with Mr 21 690,pI 7.61.Bioinformatics analysis indicated that TmTPx had a typical 2-Cys Prx conserved domain.Phylogenetic tree revealed that T.multiceps had the closest relationship to T.asiatica,followed by T.solium and T.crassiceps,E.granulosus and E.multilocularis.

关 键 词：多头带绦虫 脑多头蚴 硫氧还蛋白过氧化物酶 克隆 序列分析 

分 类 号：R345.46[医药卫生—基础医学]