Construction, identification and expression of Lactococcus lactis based EmⅡ/3 vaccine of Echinococcus multilocularis
10.3760/cma.j.cn231583-20240227-00042
- VernacularTitle:乳酸乳球菌介导的多房棘球绦虫EmⅡ/3疫苗的构建、鉴定及表达
- Author:
Wengui LI
1
;
Xingkun OU
;
Ailin HE
Author Information
1. 重庆医科大学附属第一医院传染病寄生虫病研究所，重庆 400016
- Keywords:
Echinococcus multilocularis;
Lactococcus lactis;
EmⅡ/3;
Vaccine
- From:
Chinese Journal of Endemiology
2024;43(7):517-522
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct Lactococcus lactis (LL) based EmⅡ/3 vaccine of Echinococcus multilocularis and observe its expression efficiency. Methods:EmⅡ/3 gene was obtained through PCR amplification using the pCD-EmⅡ/3 as a template, the gene was cloned into pMG36e to construct pMG36e-EmⅡ/3 and transformed into Escherichia coli BL21 (DE3) competent cell, the recombinant plasmid was identified by double restriction endonuclease digestion, then was electroporated into LL MG1363 to construct recombinant (r) LL-EmⅡ/3 vaccine. After screening for roxithromycin resistance, the plasmid was extracted for PCR identification, and the expression was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Results:The pMG36e-EmⅡ/3 recombinant plasmid was identified by double restriction endonuclease digestion, and there was an approximately 3 600 bp carrier band and a 1 759 bp EmⅡ/3 gene band. The plasmid extracted from roxithromycin resistant rLL strain was used as a template, and 1 759 bp EmⅡ/3 gene was amplified by PCR; SDS-PAGE demonstrated that the rLL-EmⅡ/3 vaccine could express EmⅡ/3 protein with a relative molecular weight of 66 × 10 3, and the expressed protein accounted for about 20% of the total bacteria protein after 3 days of culture; Western blot showed that the expressed protein could be recognized by mouse serum infected with alveolar hydatid cyst. Conclusion:We have successfully constructed the rLL-EmⅡ/3 vaccine of Echinococcus multilocularis, which expresses Em Ⅱ/3 protein with specific antigenicity.
