捻转血矛线虫Hc38基因保守结构域所在片段的克隆及原核表达  

Cloning and prokaryotic expression of Hc38 gene conserved domain of Haemonchus contorts

作　　者：赵军明[1,2] 荣光[3] 郑金海[1,2] 王新华[2] 薄新文[2] 刘志强[1,2] 黄新[1,2] 罗盘棋[1,2] 任耀军[1,2] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]新疆兵团绵羊繁育生物技术重点实验室,新疆石河子832003 [3]中国热带农业科学院,海南儋洲571737

出　　处：《中国兽医学报》2009年第2期166-169,共4页Chinese Journal of Veterinary Science

基　　金：新疆兵团科技局社会发展资助项目(2006GG31)

摘　　要：参照捻转血矛线虫Hc38基因核酸序列(登录号:AY749124)的保守结构域设计1对引物,采用RT-PCR方法从绵羊捻转血矛线虫雌性成虫中扩增出约429 bp的cDNA序列,将其克隆到pMD19-T中,经酶切鉴定证实,并进行序列测定。与AY749124序列同源性为99%。进一步将克隆基因与原核表达载体pPRO EX HTa构建重组表达载体,经IPTG诱导,能在DH5a细菌中表达了相对分子质量约17 000的融合蛋白。经Western blot等检测表明,诱导表达的抗原蛋白能与捻转血矛线虫阳性血清发生特异性反应。A pair of primers for amplifying the Hc38 gene（AYA749124） of Haemonchus contorts were densigned,with which the cDNA fragment of Hc38 was amplified from Haemonchus contorts by RT PCR and then cloned into PMD19-T vector. Sequence analysis of the fragment suggested that it possessed 99% homology with the Hc38 gene. The fragment was inserted into the expression vector pPRO EX HTa. The recombinant plasmid produced a 17 000 fusion protein in E. coli DH5a under the induction of IPTG. The expression quantity and immunoreactivity of the fusion protein was detected by SDS-PAGE electrophoresis and Western blotting.

关 键 词：捻转血矛线虫 Hc38 基因克隆 原核表达 

分 类 号：S852.731[农业科学—基础兽医学]