家畜生态学报2018，Vol.39Issue(1)：19-24,6.
细粒棘球蚴GST-mu2基因的克隆、表达及重组蛋白反应原性研究
Cloning and Expression of GST-mu2 Gene of Echinococcus granulosus and Reactionogenicity of Recombinant Protein
摘要
Abstract
In order to study the reactogenicity of glutathione s-transferase mu 2 (GST-mu2),specific primer derived from Echinococcus granulosus (Eg) genome database in GenBank was designed and the open reading frame(ORF) sequence of GST-mu2 was cloned by RT-PCR from hydatid protoscolex.Then the amplified product was cloned into pMD19-T vector,sequenced and subcloned into the expression vector pET-28a.The recombinant pET-GST-mu2 expression vector was generated successfully and then was transformed into E.coli competent cells of BL21 (DE3),induced by IPTG.Then the reactogenicity of re-combinant protein GST-mu2 was analyzed.The results showed that GST-mu2 gene had a length of 660 bp,encoding 219 amino acids containing two N-acylation sites two CK Ⅱ phosphorylation sites;three PKC phosphorylation sites;one TYR phosphorylation site.The epitopes focus on the amino acid from 28 to 70 and 147 to 219.The recombinant protein with a molecular weight of 32 kDa was examined by SDS-PAGE and further confirmed by Western blot,which displaying stronger reactionogenicity.This study laid a pre-liminary foundation for further Eg.diagnostic antigen based on GST-mu2 as a candidate protein.关键词
细粒棘球蚴/GST-mu2基因/克隆/表达/反应原性Key words
Echinococcus granulosus/GST-mu2 gene/cloning/expression/reactionogenicity分类
农业科技引用本文复制引用
陈英,乔军,孟庆玲,钟文强,刘田莉,贡莎莎,才学鹏..细粒棘球蚴GST-mu2基因的克隆、表达及重组蛋白反应原性研究[J].家畜生态学报,2018,39(1):19-24,6.基金项目
兵团国家科技合作计划(2016AH006) （2016AH006）
国家国际科技合作交流项目(CK07-11) （CK07-11）
家畜疫病病原生物学国家重点实验室开放课题(SKLVEB2016KFKT008) （SKLVEB2016KFKT008）
