本地毛形线虫HSP70基因的克隆与序列分析  

Cloning and sequence analysis of HSP70 gene from Trichinella nativa

作　　者：袁金钱[1] 路义鑫[1] 韩彩霞[1] 宋铭忻[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2007年第12期1062-1065,共4页Chinese Veterinary Science

基　　金：国家科技支撑计划重大项目(2006BAD06A09);国家自然科技资源基础平台项目(2005DKA21104);霍英东青年教师基金项目(91034)

摘　　要：根据GenBank中已发表的布氏旋毛虫HSP70基因序列设计了1对引物，用Trizol法从本地毛形线虫肌幼虫虫体中提取总RNA，用RT-PCR方法扩增了HSP70基因，将目的基因克隆至pMD18-T载体，转化大肠杆菌DH5α。重组质粒用PCR、限制性内切酶BarnHⅠ和HindⅢ进行单、双酶切鉴定。测序结果表明，成功克隆了本地毛形线虫HSP70基因。序列分析表明，HSP70基因比较保守，不同物种之间氨基酸的同源性在40％～80％。与布氏旋毛虫HSP70序列相比，CDS区多出9个碱基，但核苷酸序列和氨基酸同源性分别为98%和94％，存在1个糖基化位点和1个潜在的信号肽位点。A pair of primers for HSP70 gene of Trichinella nativa was designed according to the published HSP70 gene sequence of Trichinella britovi in GenBank （No. Y13114）. Total RNA was extracted from muscle larvae of T. nativa by Trizol. A target gene was amplified by RT-PCR and cloned into pMD18- T vector,and then transformed into Escherichia coli DH5α. The recombinant plasmids were identified by PCR and enzymatic digestion with BamHⅠ and HindⅢ. Sequencing result showed that HSP70 gene of T. nativa was cloned successfully. Sequence analysis indicated that HSP70 gene was relatively conservative, and homology of amino acids among different species was 40%- 80%. In the CDS region of the amplified HSP70 gene,there were 9 bases more than that of T. britovi,and the homology of nucleotide sequence and amino acids was 98% and 94% between T. nativa and T. britovi,respectively. There were one potential glycosylation site and one signal peptide site in the deduced amino acid sequence of the HSP70 gene of T. nativa.

关 键 词：本地毛形线虫 HSP70基因 序列分析 

分 类 号：S852.731[农业科学—基础兽医学]