Journal article
Characterization of caprine interleukin-4
Veterinary immunology and immunopathology, Vol.78(3), pp.219-229
2001
Handle:
https://hdl.handle.net/2376/109002
PMID: 11292525
Abstract
Caprine interleukin-4 (IL-4) cDNA was cloned from RNA of mitogen stimulated goat peripheral blood mononuclear cells utilizing reverse transcriptase–polymerase chain reaction. The sequence of caprine IL-4 cDNA corresponds to a 535 nucleotide mRNA with 5′- and 3′-untranslated regions and a 405 nucleotide open reading frame, the first 66 nucleotides of which encode a putative signal peptide. Mature IL-4 is a 12.8
kDa protein containing six cysteine residues and two potential
N-linked glycosylation sites and is highly homologous with other ruminant IL-4. The predicted molecular mass of mature unglycosylated IL-4 was confirmed by western blot of recombinant caprine IL-4 expressed in bacteria with a monoclonal antibody against a carboxyterminal peptide derived from the predicted amino acid sequence of bovine IL-4. Eukaryotic expression plasmids containing caprine IL-4 cDNA were used to characterize recombinant IL-4. Transcription of IL-4 mRNA was confirmed by transfection of COS-7 and goat synovial membrane cells, and recombinant IL-4 produced by stably transfected L929 cells inhibited inducible nitric oxide synthase in macrophages. Genetic immunization of mice with a caprine IL-4 cDNA expression plasmid induced antibodies against recombinant caprine IL-4 produced in bacteria.
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Details
- Title
- Characterization of caprine interleukin-4
- Creators
- K.R Snekvik - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USAJ.C Beyer - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USAG Bertoni - Institute of Veterinary Virology, University of Berne, Berne, SwitzerlandB.R Von Beust - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USAT.V Baszler - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USAG.H Palmer - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USAT.F McElwain - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USAW.P Cheevers - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USA
- Publication Details
- Veterinary immunology and immunopathology, Vol.78(3), pp.219-229
- Academic Unit
- Veterinary Microbiology and Pathology, Department of; Paul G. Allen School for Global Animal Health
- Publisher
- Elsevier B.V
- Identifiers
- 99900547209701842
- Language
- English
- Resource Type
- Journal article