Expression and biological activity of porcine interleukin-18 in recombinant Lactococcus lactis.
- Author:
Lu MA
;
Xinyuan QIAO
;
Lijie TANG
;
Yanping JIANG
;
Wen CUI
;
Yijing LI
- Publication Type:Journal Article
- MeSH:
Animals;
Blotting, Western;
Electrophoresis, Polyacrylamide Gel;
Electroporation;
Genetic Vectors;
Interleukin-18;
biosynthesis;
Lactococcus lactis;
metabolism;
Reverse Transcriptase Polymerase Chain Reaction;
Swine
- From:
Chinese Journal of Biotechnology
2014;30(10):1541-1548
- CountryChina
- Language:Chinese
-
Abstract:
To obtain active protein of pIL-18 expression in Lactococcus lactis, and to observe its biological activity, the total RNA was extracted as template from peripheral blood mononuclear cells. Porcine interleukin 18 (pIL-18) was amplified by RT-PCR. The resulting fragment was cloned into pAMJ399 L. lactis vector, and then transformed to L. lactis MG1363 cells by electroporation. Expression of pIL-18 protein was detected by SDS-PAGE and Western-blotting. Bioactivity of the product was tested by pig spleen lymphocyte proliferation test and cytopathogenic effect inhibition assay. The result of Western blotting and bioactivity test shows that the molecular weight of pIL-18 protein was 19 kDa. The react line was observed in both supernatant and precipitated of the recombinant bacteria pAMJ399-pIL18/MG1363. The expressed pIL-18 can promote the proliferation of pig spleen lymphocyte, and significantly inhibit virus multiplication. As conclusion, porcine interleukin-18 was successfully expressed in L. lactis, and the product was biologically active.
