Construction of lentivirus-mediated stable expression of duck interferon-γ cell line
Interferon-γ is one of the cytokines with antiviral activity and immunomodulatory ability.To construct a cell line stably expressing duck interferon-γ,the gene of duck interferon gamma(dIFN-γ)was optimized referring to mouse codon and synthesized.Synthesed dIFN-γ was cloned into lentiviral expressing vector plenti-GIII-CMV-GFP-2A-Puro to construct recombinant plasmid plenti-IFN.The recombinant lentiviral rlenti-IFN was rescued by co-transfected plenti-IFN and helper plasmid into packaging cell 293T.After infection Chinese hamster ovary(CHO)cells with recombiant rlenti-IFN,30 monoclonal cell lines with purinomycin resistance were selected by limited dilution and pressure screening.At last,one monoclonal cell line with the highest duIFN-γ mRNA level was selected by RT-qPCR method.One stable cell line was finally obtained by expanding culture.Western blot analysis indicated that duIFN-γ was stably expressed in this cell line.This study lays a foundation for studying duIFN-γ biological function,establishing duIFN-γ detection method and producing cheap interferon for duck or poultry.
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万方数据
维普
