Cloning,expression and immunity analysis of transketolase of Echinococcus granulosus
10.16250/j.32.1374.2017229
- VernacularTitle:细粒棘球绦虫转酮醇酶克隆表达及免疫性分析
- Author:
De-Ping CAO
1
;
Yao-Gang ZHANG
;
Chao-Qun LI
;
Jia LIU
;
Bo-Fan JIANG
Author Information
1. 青海大学医学院病原生物学教研室 西宁810001;青海大学附属医院、青海省包虫病研究重点实验室
- Keywords:
Echinococcus granulosus;
Echinococcosis;
Transketolase;
Recombinant plasmid;
Diagnostic antigen
- From:
Chinese Journal of Schistosomiasis Control
2018;30(2):155-160
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain the prokaryotic expression of transketolase genes and analyze its value as a diagnostic anti-gen for echinococcosis.Methods TK gene was amplified by PCR and cloned into prokaryotic vector pMD19-EgTK,and then subcloned into the expression vector pET-28a.The target gene TK prokaryotic expression plasmid pET-28a was constructed and transferred into BL21. The purified protein was identified by SDS-PAGE and Western blotting. The blood samples of patients with cystic echinococcosis(CE group),alveolar echinococcosis(AE group)and healthy people(healthy group)were collected and detected by ELISA with the recombinant EgTK protein as a diagnostic antigen.Results The recombinant plasmid pET-28a (+)-EgTK was constructed successfully,and there was a band around 70 kDa by using Western blotting.ELISA showed that the difference among the 3 groups of sera reaction A450was significantly different(F=44.47,P<0.01),and the A450values of the CE group(1.46±0.41)and AE group(1.28±0.29)were higher than that of the healthy group(0.66 ± 0.23),but there was no significant difference between the former two.Conclusion The recombinant EgTK protein is better to distinguish the echinococ-cosis group and healthy group,but it can't do a differential diagnosis between CE and AE cases.
