First Author | Muneta Y | Year | 2000 |
Journal | Cytokine | Volume | 12 |
Issue | 6 | Pages | 566-72 |
PubMed ID | 10843730 | Mgi Jnum | J:63548 |
Mgi Id | MGI:1861163 | Doi | 10.1006/cyto.1999.0648 |
Citation | Muneta Y, et al. (2000) PORCINE INTERLEUKIN 18: CLONING, CHARACTERIZATION OF THE cDNA AND EXPRESSION WITH THE BACULOVIRUS SYSTEM. Cytokine 12(6):566-72 |
abstractText | We have isolated and sequenced a cDNA that contains the coding sequence of porcine interleukin 18 (IL-18) and the recombinant protein of porcine IL-18 was expressed using the baculovirus system. The open reading frame (ORF) of the porcine IL-18 cDNA is 579 base pairs (bp) in length and encodes 192 amino acids. The predicted amino acid sequence is 76.7%, 64.7% and 61.6% homologous to the predicted human, murine and rat amino acid sequences, respectively. The porcine precursor and mature IL-18 protein were expressed respectively in Trichoplusia ni -derived (Tn5) cells using the baculovirus Autografha californica nuclear polyhedorosis virus (AcNPV) as a vector. Tn5 cells infected with recombinant virus containing a whole IL-18 protein coding region sequence secreted porcine precursor IL-18 into the culture medium. On the other hand, Tn5 cells infected with recombinant virus containing a mature IL-18 protein coding region sequence expressed several proteins in the cell lysates, but did not secrete mature protein into the culture medium efficiently. Immunoblotting analysis of recombinant protein showed cross-reactivity with anti-human IL-18 polyclonal antibody. The mature form of porcine IL-18 protein induced IFN-gamma production in suboptimal doses of anti-CD3 antibody and concanavalin A- (ConA) stimulated porcine peripheral blood mononuclear cells (PBMC), but the precursor form had little effect. Copyright 2000 Academic Press. |