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  • Title: [Eukaryotic expression of recombinant porcine single-chain IL-12 and its biologic activity].
    Author: Wang XM, Yuan YY, Fang Q, Xia H, Sun X, Li BQ.
    Journal: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi; 2012 Jul; 28(7):691-5. PubMed ID: 22768856.
    Abstract:
    AIM: To express the recombinant porcine single-chain interleukin-12 (pscIL-12) gene in CHO-K1 cells, and identify biological activity of pscIL-12 fusion protein. METHODS: The recombinant pcDNA3.1(+)-pscIL-12 plasmid was transfected into the CHO-K1 cells using Sofast(TM); reagent. The levels of pscIL-12 fusion protein and IFN-γ produced by human peripheral blood mononuclear cells (PBMCs) stimulated with the supernatant of CHO-K1 cells were detected by ELISA. NK cell cytotoxicity was tested by MTT assay. The lymphocyte proliferation was examined by flow cytometry. RESULTS: ELISA showed that the transfected CHO-K1 cells had a relatively higher expression of pscIL-12 fusion protein. And the pscIL-12 fusion protein possessed strong bioactivities in enhancing the NK cell cytotoxicity, increasing the IFN-γ production, and stimulating the lymphocyte proliferation. CONCLUSION: CHO-K1 cells transfected with pcDNA3.1(+)-pscIL-12 recombinant plasmid can successfully express pscIL-12 fusion protein, which has the expected biological activities.
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