畜牧与兽医2018，Vol.50Issue(3)：63-67,5.
细粒棘球蚴EG19抗原基因的克隆表达及重组蛋白反应原性研究
Cloning and expression of EG19 gene of Echinococcus granulosus and reactionogenicity of recombinant protein
摘要
Abstract
In order to study the reactionogenicity of Eg19 gene,primers derived from the Echinococcus granulosus (Eg) genome database in GenBank were designed and the open reading frame (ORF) sequence of Eg19 was amplified by RT-PCR from hydatid protoscolex.Then the amplified products were cloned into the pMD19-T vector,sequenced and subeloned into the expression vector pET-32a.The recombinant pET-Eg19 expression vector was generated successfully and then was transformed into the E.coli competent cells of BL21 (DE3),induced by IPTG.Finally,the reactionogenicity of the recombinant protein Eg19 was analyzed.The results showed that Eg19 had a length of 534 bp,encoding 177 amino acids that contained two N-myristoylation sites,one CK Ⅱ phosphorylation site,one PKC phosphorylation site and one ATP/GTP-binding site motif A (P-loop).The epitope regions are concentrated on the amino acids from 20 to 93 and 96 to 146.The recombinant protein with a molecular weight of 35 kDa could be identified by SDS-PAGE and was confirmed by Western blot,which displayed strong reactionogenicity.This study laid a preliminary foundation for diagnosis of E.g.infection using Eg19 as a candidate antigen.关键词
细粒棘球蚴/Eg19抗原基因/克隆/表达/反应原性Key words
Echinococcus granulosus/Eg19/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,王熙凤,黄运福,才学鹏..细粒棘球蚴EG19抗原基因的克隆表达及重组蛋白反应原性研究[J].畜牧与兽医,2018,50(3):63-67,5.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）
