Molecular cloning and expression of cynomolgus monkey interferon-gamma cDNA.
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.
Tatsumi
M
M
Department of Veterinary Science, AIDS Research Center, National Institute of Infectious Diseases, Tokyo, Japan.
Sata
T
T
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D003208:Concanavalin A / Q000494:pharmacology; D003588:Cytopathogenic Effect, Viral; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics* / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects; D008252:Macaca fascicularis / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses; D016133:Polymerase Chain Reaction; D051381:Rats; D011994:Recombinant Proteins; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D012756:Sheep; D018411:Spodoptera / Q000821:virology; D013552:Swine
D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D003208:Concanavalin A; D007371:Interferon-gamma