中国人兽共患病学报2017，Vol.33Issue(11)：967-971,5.DOI:10.3969/j.issn.1002-2694.2017.11.004
多房棘球蚴LDH基因的克隆表达及免疫原性研究
Cloning and prokaryotic expression of LDH gene in Echinococcus multilocularis and immunogenicity research of the recombinant protein
摘要
Abstract
The present study aimed to clone and express the EmLDH gene of Echinococcus multilocularis in Qinghai Province,identifying immunogenicity of EmLDH recombinant protein and evaluating its immune diagnostic value preliminarily.EmLDH genes were cloned by RT-PCR technology and linked into pET15b vector.Recombinant expression pET15b-EmLDH vectors were constructed and transformed into E.coli Rosetta (DE3) competent cells.Recombinant proteins were induced and expressed.Expression forms of recombinant proteins were detected by SDS-PAGE.Recombinant proteins were purified by affinity chromatography of Ni-IDA resin.Immunogenicity of recombinant proteins was identified by Western blotting.Serum samples from patients with alveolar echinococcosis (57 cases),cystic echinococcosis (33 cases),and healthy persons (50 cases) were examined by ELISA,which evaluated preliminarily immune diagnosis effect of EmLDH recombinant proteins.Results showed that EmLDH gene was cloned successfully and the recombinant proteins were expressed and purified.Results of Western blotting showed EmLDH recombinant proteins were recognised by serum samples from patients with alveolar echinococcosis and cystic echinococcosis,but not by serum samples from healthy persons.Results of ELISA showed that diagnostic sensitivities of EmLDH recombinant protein reacted with serum samples from patients with alveolar echinococcosis and cystic echinococcosis were 84.21％ and 84.85 ％ respectively.EmLDH recombinant proteins of Echinococcus multilocularis have high immunogenicity and good immune diagnostic value for echinococcosis.关键词
多房棘球蚴/LDH基因/克隆/表达/免疫原性Key words
Echinococcus multilocularis/LDH gene/cloning/expression/immunogenicity分类
医药卫生引用本文复制引用
何顺伟,李洪清,李晓燕,赵瑞雪,魏晓星..多房棘球蚴LDH基因的克隆表达及免疫原性研究[J].中国人兽共患病学报,2017,33(11):967-971,5.基金项目
青海省科技项目(No.2016-ZJ-746)资助 Supported by the Science and Technology Project of Qinghai Province (No.2016-ZJ-746) （No.2016-ZJ-746）
