Cloning, Transformation and Expression of Human IFN-γ by Genetic Engineering in Saccharomyces cerevisiae

Abstract

Human interferon-gamma (hIFN-γ) is a Glycoprotein pertinence to a distinct group of interferon, called type II interferons, which have an immunological function to respond to antigenic stimuli such as bacteria, viruses, fungi, and any infections by a microorganism. The hIFN-γ is produced by natural killer T (NKT) and natural killer (NK) cells during the immune response as part of the innate immune response and by Th1 CD4 and CD8 cytotoxic T lymphocytes (CTL) effector T cells upon the development of antigen-specific immunity. Recombinant hIFN-γ has been produced in different expression systems comprising prokaryotic, insect, fungal (yeasts), protozoan, mammalian cells, and plant. In the present study, pET28a plasmid was used in this research for preparation to insert and pYES2 plasmid for cloning and expression of human IFN-γ gene in yeast. The trademark of hIFN-γ has been produced in Escherichia coli is termed ACTIMMUNE®, but the human interferon- gamma was produced in the prokaryotic expression system is unglycosylated form. This result increases in cost due to increased purification, as well as short-life in the bloodstream, but it is biologically active. This study aimed to use Saccharomyces cerevisiae 4741 strain as a eukaryotic system for expression of hIFN-γ cDNA instead of a prokaryotic system in glycosylation patterns as modified translation. But the results are not satisfactory as the produce of yields.