EXPRESSION AND CHARACTERIZATION OF A FUSION PROTEIN TS-HSP70 AND TS87 OF TRICHINELLA SPIRALIS
10.3969/j.issn.1005-0507.2015.01.001
- VernacularTitle:旋毛虫Hsp70与Ts87融合蛋白的构建表达及鉴定
- Author:
Liang SUN
1
;
Qing SUN
;
Lei FANG
;
Jing YANG
;
Yuan GU
;
Kuo BI
;
Xin-Ping ZHU
Author Information
1. 首都医科大学基础医学院人体寄生虫学教研室
- Keywords:
Trichinella spiralis;
Recombinant fusion protein;
Heat shock protein 70(HSP70);
Excretory-secretory antigen;
Vaccine
- From:
Acta Parasitologica et Medica Entomologica Sinica
2015;(1):1-6
- CountryChina
- Language:Chinese
-
Abstract:
To construct and express the fusion protein that combines Trichinella spiralis heat shock protein 70 with excretory-secretory antigen Ts87, restriction enzyme EcoRⅠ site was applied to fuse Ts-Hsp70 with Ts87.The target genes were inserted into plasmid pET28-a ( +), and transformed into E.coli.BL21 (DE3).After expression and purification, the fusion protein rTs-Hsp70-Ts87 was characterized by SDS-PAGE and Western blot.The results showed that the fusion gene was 2 721 bp.The results of PCR, enzyme digestion and sequencing analysis showed that the recombinant plasmid was affirmed to be successfully constructed.The fusion protein rTs-Hsp70-Ts87 was expressed with IPTG induction and purified by Ni-affinity chromatography and refolded.The results of SDS-PAGE and Western blot indicated that molecular weight of the protein is about 100 kDa, and it could be recognized by anti His-tag monoclonal antibody, anti-rTs-Hsp70 immune serum and anti-rTs87 immune serum.In conclusion, T.spiralis recombinant fusion protein rTs-Hsp70-Ts87 was constructed and expressed successfully, which had laid the groundwork for further research.
