Preparation and characterization of a recombinant poly-epitopic vaccine EgG1Y162-2 (4) against cystic echinococcosis based on the linker GSGGSG
10.16250/j.32.1374.2022116
- VernacularTitle:基于接头序列“GSGGSG”的细粒棘球蚴病重组 多表位疫苗EgG1Y162-2 (4) 的制备及鉴定
- Author:
Jia ZHENG
1
;
Dong-jun ZHANG
2
;
Shang-qi ZHAO
1
;
Yan-min LI
1
;
Yan-xia ZHOU
1
;
Wen-tao ZHOU
3
;
Xiao-tao ZHOU
1
Author Information
1. Department of Immunology, School of Basic Medicine, Xinjiang Medical University, Urumqi, Xinjiang 830054, China
2. Shandong Institute of Parasitic Disease Control, Shandong First Medical University, Jining, Shandong Province, China
3. The Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang Uygur Autonomous Region, China
- Publication Type:Journal Article
- Keywords:
Cystic echinococcosis;
Linker sequence;
Multi-epitope vaccine;
EgG1Y162-2 (4);
Prokaryotic expression;
Characterization
- From:
Chinese Journal of Schistosomiasis Control
2022;34(4):378-382
- CountryChina
- Language:Chinese
-
Abstract:
Objective To perform prokaryotic expression and preliminary characterization of the recombinant poly-epitope vaccine EgG1Y162-2 (4) against cystic echinococcosis. Methods The recombinant poly-epitope vaccine EgG1Y162-2 (4) against Echinococcus granulosus based on the linker GSGGSG was subjected to structural three-dimensional (3D) modeling using immunoinformatics to analyze the structural changes and evaluate the antigenicity of the vaccine. The pET30a-EgG1Y162-2 (4) recombinant plasmid was generated using double digestion with EcoR I and Sal I, and then transformed into competent cells. Following protein induction with isopropyl-β-D-thiogalactoside (IPTG), the prokaryotic expression proteins were characterized using Western blotting, and the antigenicity of the recombinant protein was analyzed using sera from cystic echinococcosis patients and health volunteers. Results The four EgG1Y162-2 proteins coupled by the 3D structure of the recombinant vaccine EgG1Y162-2 (4) presented independent and effective expression and good antigenicity. The highest protein expression was detected in the supernatant following induction of the recombinant plasmid pET30a-EgG1Y162-2 (4) by 0.2 mmol/L IPTG at 37 °C for 4 h, and a pure protein component was seen following elution with 60 mmol/L imidazole. Western blotting analysis of the recombinant multiepitope protein HIS-EgG1Y162-2 (4) showed a band at approximately 39 kDa, and this band was recognized by sera from cystic echinococcosis patients. Conclusion A recombinant poly-epitope vaccine EgG1Y162-2 (4) against cystic echinococcosis has been successfully constructed, which provides a preliminary basis for researches on recombinant multi-epitope vaccine against cystic echinococcosis.
