豆状带绦虫胰岛素样生长因子受体基因的克隆分析及原核表达  

Cloning,bioinformatics analysis and prokaryotic expression of insulinlike growth factor receptor gene from Taenia pisiformis

作　　者：杨锐[1] 张少华[2] 骆学农[2] 史云洁 曾巧英[1] 才学鹏[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病病原生物学国家重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2016年第4期403-408,共6页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31372433);甘肃省创新研究群体计划项目(1210RJIA006)

摘　　要：采用RT-PCR和cDNA末端快速扩增(RACE)法克隆了豆状带绦虫(Taenia pisiformis)胰岛素样生长因子受体基因(TpIGFR)的全长cDNA序列,并对其进行了生物信息学分析和原核表达。结果显示,Tp IGFR cDNA全长5 154 bp,其ORF为4 953 bp,编码1 651个氨基酸。系统进化分析表明,TpIGFR与多房棘球绦虫、猪带绦虫胰岛素样生长因子受体的相似性均大于83%,处于同一进化分支。蛋白结构预测表明,该蛋白由胞外配体结合区、跨膜区和酪氨酸蛋白激酶区组成,具有IGFR的特征结构域。Western-blot结果表明,该重组蛋白能被豆状囊尾蚴感染家兔血清及抗His抗体特异识别,具有反应原性。本研究为后续带科绦虫IGFR基因的功能研究奠定了基础。A full-length c DNA encoding Taenia pisiformis insulin-like growth factor receptor（TpIGFR） gene was obtained by RT-PCR and rapid amplification of c DNA ends methods. Sequence analysis showed that the c DNA sequence obtained was 5 154 bp in length with a 4 953 bp open reading frame encoding 1 651 amino acids.The phylogenetic analysis showed that TpIGFR gene had 83% homology with that of Echinococcus multilocularis and Taenia solium,and these three genes were classified into the same branch. The predication of protein structure revealed that TpIGFR had the typical structural domain of IGFR,including extracellular ligand-binding domain,transmembrane domain and tyrosine protein kinase domain. Western-blotting confirmed that the Escherichia coli-expressed recombinant protein p ET-30a（＋）-TpIGFR-LD was specifically recognized by anti-His antibody and showed a positive reaction with rabbit positive serum sample infected with T. pisiformis metacestode,indicating that recombinant TpIGFR-LD is antigenic.This study is valuable in studying the IGFR functions of tapeworm in the future.

关 键 词：豆状带绦虫 RACE TpIGFR基因 原核表达 

分 类 号：S852.734[农业科学—基础兽医学]