Characterization of caprine interleukin-4

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 nucleot...

Full description

Saved in:
Bibliographic Details
Published in:Veterinary immunology and immunopathology 2001-02, Vol.78 (3), p.219-229
Main Authors: Snekvik, K.R., Beyer, J.C., Bertoni, G., Von Beust, B.R., Baszler, T.V., Palmer, G.H., McElwain, T.F., Cheevers, W.P.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibodies, Monoclonal - immunology
Base Sequence
Blotting, Western - veterinary
Caprine
Cattle
Cloning, Molecular
COS Cells
DNA, Complementary - chemistry
Expression plasmids
Goats - genetics
Goats - metabolism
iNOS assay
Interleukin-4
Interleukin-4 - genetics
Interleukin-4 - immunology
Mice
Molecular Sequence Data
Molecular Weight
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Transfection
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0165-2427
1873-2534
DOI:10.1016/S0165-2427(01)00233-1