Cloning and expression of human IFN-γ in Leishmania tarentolae

Autor: Mahdi Hemayatkar, Azam Hemmati, Ahmad Adeli, Noushin Davoudi, Zahra Khodayari
Rok vydání: 2011
Předmět:
Zdroj: World Journal of Microbiology and Biotechnology. 27:1893-1899
ISSN: 1573-0972
0959-3993
DOI: 10.1007/s11274-010-0648-4
Popis: Increasing therapeutic applications for recombinant human interferon-gamma (rhIFN-γ), 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-γ cDNA was amplified from phyto-hemagglutinin-stimulated peripheral blood mononuclear cells of a healthy blood donor using RT–PCR. In order to express, the rhIFN-γ protein, the resulting cDNA was cloned in two expression cassettes (each containing one copy of hIFN-γ 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-γ and ELISA confirmed the expression and production of 9.5 mg of rhIFN-γ protein/l respectively.
Databáze: OpenAIRE
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