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A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
Published on: August 31, 2014
Molecular cloning and expression of cynomolgus monkey interferon-gamma cDNA
1Department of Veterinary Science, AIDS Research Center, National Institute of Infectious Diseases, Tokyo, Japan.
Abstract:
Since the discovery of its involvement in the pathogenesis of simian immunodeficiency virus infection, the role of macaque monkey interferon-gamma (IFNgamma) has been a focus of particular interest. The purpose of this study was to express bioactive recombinant monkey IFNgamma, as well as clone cynomolgus monkey IFNgamma cDNA. The isolation of a cDNA encoding cynomolgus monkey IFNgamma was performed using a reverse transcriptase polymerase chain reaction-based strategy with activated monkey lymphocyte cDNA as templates. Cynomolgus monkey IFNgamma consists of 165 amino acids including a putative signal sequence and has 94, 60 and 39% identity to human, bovine and murine homolgues, respectively. Monkey IFNgamma cDNA was highly expressed as three distinct secreted proteins by a recombinant baculovirus system. The secreted proteins were revealed to have antiviral activity up to 10(8) units/ml characteristic of authentic IFNgamma by a bioassay of a cytopathic effect reduction assay.
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