Increasing therapeutic applications for recombinant human interferon-gamma (rhIFN-c), an antiviral pro-inflammatory cytokine, has broadened interest in optimizing methods for its production. We herein describe a unicellular eukaryotic system, Leishmania tarentolae, a Trypanosomatidae protozoan parasite of gecko Tarentola annularis, which has recently been introduced as a candidate for heterologous gene expression. In this study, the hIFN-c cDNA was amplified from phyto-hemagglutininstimulated peripheral blood mononuclear cells of a healthy blood donor using RT-PCR. In order to express, the rhIFNc protein, the resulting cDNA was cloned in two expression cassettes (each containing one copy of hIFN-c cDNA) and integrated into the small subunit of ribosomal RNA gene of L. tarentolae genome by electroporation. Transformed clones were selected in the presence of appropriate antibiotics. Western blotting of rhIFN-c and ELISA confirmed the expression and production of 9.5 mg of rhIFN-c protein/l respectively.
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