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  • Tetracycline-controlled expression of glycosylated porcine interferon-gamma in mammalian cells
    Cencič, Avrelija ...
    Tetracyclinne-controlled expression plasmids that aloww inducible expression of proteins in mammalian cells (Gossen & Bujard, 1992), have been used to express porcine interferon-gamma in RK-13 rabbit ... kidney cell line. Following neomycin selection, stable clones produced recombinant, glycosylated porcine interferon-gamma only after removal of tetracycline (Tc). Southern blot analysis of one clone showed that approximately 50 copies of IFN-gamma cDNA were present in the cell genome. In the absence of Tc, stable clones secreted large amounts of rGPoINF-gamma (up to 16 ug/ml) into the medium supplemented with 10% FCS and high glucose concentration. Molecular weight comparison of s-Methionine, labelled rGPoIFN-gamma with natural leukocytic IFN-gamma aftr immunoprecipitation, revealed 4 major glycoforms with apparent Mr of 27,000; 25,000; 20,000 and 18,500, that are almost identical in both IFN-gamma species. In both cases, all 4 glycoforms resolved into 2 polypeptide monomers with apparent Mr of 16,500 and 14,500 upon deglycosylation with N-glycosydase F. The biological activity of rGPoINF-gamma was in the same range as that of natural leukocytic PoINF-gamma. Eventually, this recombinant mammalian IFN-gamma should constitute a very useful substitute for leukocyte PoINF-gammain vitro in in vivo experiments.
    Vir: Animal biotechnology. - ISSN 1049-5398 (Let.10, št. 1/2, 1999, str. 63-79)
    Vrsta gradiva - članek, sestavni del
    Leto - 1999
    Jezik - angleški
    COBISS.SI-ID - 1615148

vir: Animal biotechnology. - ISSN 1049-5398 (Let.10, št. 1/2, 1999, str. 63-79)
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