弓形虫棒状蛋白17基因的克隆与序列分析  

Cloning and Sequence Analysis of ROP17 Gene of Toxoplasma gondii

作　　者：聂子雄[1,2] 张念章[2] 胡钱江[1] 陈锐钊[1,2] 周东辉[2] 袁子国[1] 林瑞庆[1] 朱兴全[2] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [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,甘肃兰州730046

出　　处：《中国畜牧兽医》2013年第4期13-17,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(31230073、31172316、31101812);全国农业科研杰出人才项目;中央级公益性科研院所基本科研业务费专项(2012ZL081);中国农业科学院院长基金项目

摘　　要：为研究弓形虫棒状体蛋白17(ROP17)基因的遗传变异,本研究以弓形虫RH株、PRU株和TgC7株为研究对象,首次PCR扩增了其ROP17基因部分序列,将PCR产物纯化后克隆到pMD18-T并测序。将测定的序列与网上下载的弓形虫VEG株、GT1株和ME49株相应序列进行比对,然后用Mega 5.0程序的NJ法和Puzzle 5.2程序的ML法构建系统发育树。结果表明,3株弓形虫分离株的ROP17基因部分序列长度均为1375bp,A+T含量在49.53%～50.04%之间,3个虫株相应序列的变异碱基数皆小于20个,其变异率为2.3%,氨基酸序列变异率在0～6.11%之间。系统发育结果显示,ROP17基因部分序列能区分弓形虫基因Ⅰ型和Ⅱ型的虫株。本研究结果为进一步研究弓形虫ROP17基因的变异及研制弓形虫ROP17基因的亚单位疫苗提供了理论依据。Toxoplasma gondii ROP17 gene was amplified from individual T.gondii strains from different geographical locations and hosts by PCR. These PCR products were subjected to be cloned and sequenced and ROP17 sequences were aligned using the ClustalX 1.81. The phylogenetic relationships among T.gondii strains were constructed using the software Mega 5.0 and Puzzle 5.2. The lengths of all ROP17 sequences were 1375 bp, and the A＋T contents were 49.53% to 50.04%. The intra-specific variation among the T.gondii strains was up to 2.23%. The variation rate in amino acid sequences was 0 to 6.11%. Phylogenetic analysis revealed that ROP17 gene sequences could be used as a marker for studying genetic relationships of T.gondii isolates. The low variation in ROP17 gene sequences among T.gondii strains might indicate that ROP17 gene could be used as a potential anti-toxoplasmosis vaccine candidate molecule in further studies.

关 键 词：刚地弓形虫 ROP17基因 克隆 序列分析 

分 类 号：Q78[生物学—分子生物学]