猪带绦虫烯醇化酶基因的原核表达及鉴定  

Prokaryotic expression and identification of enolase gene fromTaenia solium

作　　者：尤亚南 李继东[2,3] 骆学农[2] 张少华[2] 苏君鸿[2] 李雪强[2] 侯俊玲[2] 岳城[1] 才学鹏[2] 

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830052 [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农业部兽医公共卫生重点实验室甘肃省动物寄生虫病重点实验室,甘肃兰州730046 [3]宁夏大学农学院,宁夏银川750021

出　　处：《中国兽医科学》2014年第3期221-226,共6页Chinese Veterinary Science

基　　金：甘肃省自然科学基金项目(1010RJZA002);甘肃省创新研究群体计划项目(1210RJIA006)

摘　　要：为了探索猪带绦虫烯醇化酶基因的生物学功能,以猪带绦虫幼虫cDNA为模板,PCR扩增获得烯醇化酶基因的ORF,并将其克隆至原核表达载体pET-30a(+),构建重组表达质粒。经IPTG诱导表达后,纯化表达产物,采用免疫印迹法分析表达产物的免疫反应性,并测定它与纤溶酶原的结合特性及酶活力。结果显示,该基因的ORF大小为1 302bp,表达大小约为53ku的重组蛋白;表达产物可被猪囊尾蚴病阳性血清识别;该重组蛋白具有纤溶酶原结合特性及酶活力。据此推测,猪带绦虫烯醇化酶可能在寄生虫入侵宿主过程中发挥作用,有作为药物靶标或疫苗候选分子的潜力。In order to explore the biological function of enolase of Taenia solium,its ORF was ampli- fied from larvae by RT-PCR and subeloned into the prokaryotic expression vector pET-30a （＋） to con- struct recombinant plasmid. The expressed protein induced by IPTG was purified and its immunoreactivity was examined by Western-blot. The interaction of the recombinant protein with plasminogen,and the en- zyme activities of the recombinant protein were also determined. In result,the size of ORF was 1 302 hp. The protein encoded by the enolase gene was 53 ku in size and was recognized by the serum of swine infec- ted with T. solium metacestodes,and it also had the plasminogen binding property and enzyme activities. It is proposed that enolase of T. solium plays a role in invading hosts,and has a potential use as a drug target or vaccine candidate.

关 键 词：猪带绦虫 烯醇化酶基因 表达 纤溶酶原 酶活力 

分 类 号：S852.734[农业科学—基础兽医学]