长角血蜱MLC基因的原核表达及表达产物的免疫原性分析  

Prokaryotic expression of MLC gene of Haemaphysalis longicornis and immunogenicity analysis of the recombinant protein

作　　者：田美媛[1] 田占成[2] 谢俊仁[2] 罗金[2] 沈辉[2] 袁小松[2] 郑进峰[2] 王芳芳[2] 曾巧英[1] 刘光远[1,2] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国农业科学院兰州兽医研究所家畜疫原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：《中国兽医科学》2012年第12期1211-1215,共5页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31201899)

摘　　要：为了解长角血蜱(Haemaphysalis longicornis)肌球蛋白碱轻链(MLC)基因表达产物的免疫学活性,利用RT-PCR方法扩增了长角血蜱MLC基因的完整开放阅读框,将该基因连接到原核表达载体pGEX-4T-1中进行表达,在37℃、1.0mmol/L IPTG诱导条件下,用SDS-PAGE分析目的蛋白的表达情况。结果表明,该重组蛋白的表达分子质量为44ku。Western-blotting结果显示,MLC蛋白能被兔抗长角血蜱成蜱阳性血清识别,表明MLC蛋白具有良好的反应原性。To understand the immunological characteristics of MLC gene from Haemaphysalis longicornis,the open reading frame（ORF） of MLC gene was amplified by RT-PCR,cloned into pGEX-4T-1 vector and expressed in Escherichia coli BL21（DE3）at 37 ℃ under 1.0 mmol/L IPTG induction condition.The expression of recombinant protein was analyzed by SDS-PAGE.It was found that the recombinant protein was about 44 ku in size.The MLC protein could be recognized by the positive serum against H.Longicornis adult ticks in Western-blot,which suggests that the MLC protein has immunogenicity.The study provided the basis for exploring MLC protein physiological function and for screening anti-tick vaccine candidate.

关 键 词：长角血蜱 肌球蛋白碱轻链基因 原核表达 免疫原性 

分 类 号：S852.746[农业科学—基础兽医学]