Cloning and prokaryotic expression of LDH gene in Echinococcus multilocularis and immunogenicity research of the recombinant protein
10.3969/j.issn.1002-2694.2017.11.004
- VernacularTitle:多房棘球蚴LDH基因的克隆表达及免疫原性研究
- Author:
wei Shun HE
1
;
qing Hong LI
;
yan Xiao LI
;
xue Rui ZHAO
;
xing Xiao WEI
Author Information
1. 青海大学生态环境工程学院
- Keywords:
Echinococcus multilocularis;
LDH gene;
cloning;
expression;
immunogenicity
- From:
Chinese Journal of Zoonoses
2017;33(11):967-971
- CountryChina
- Language:Chinese
-
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.
