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ARS Home » Research » Publications at this Location » Publication #96968

Title: GENE CLONING AND PROTEIN EXPRESSION OF CHICKEN IL-15

Author
item Choi, Kang
item Lillehoj, Hyun

Submitted to: Plant and Animal Genome Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 12/6/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: A cDNA encoding chicken interleukin-15 was cloned from a CD4 T cell hybridoma expression library by screening with a rabbit antibody against a protein fraction of conditioned medium containing T cell growth promoting activity. The chicken IL-15 cDNA contains an open reading frame of 143 amino acids with a single potential N-linked glycosylation site. The predicted m.w. of the encoded protein (16 kDa) matched the size of an immunoreactive band on Western blots of E. coli expressing the recombinant IL-15. Amino acid and nucleotide sequence analyses of chicken IL-15 revealed 34% and 46% homology with bovine IL-15 respectively and lesser homologies to other mammalian IL-15s. present in mammalian IL-15 sequences. RT-PCR demonstrated that the chicken IL-15 gene is expressed in many tissues including spleen, intestine, and muscle and in established macrophage, T lymphoma and fibroblast cell lines. Activation of spleen cells with Con A enhanced the expression of IL-15 gene transcripts in a time-dependent manner. CHO-K1 cells transfected with the chicken IL-15 cDNA secreted a biologically active protein supporting the growth of Con A activated spleen lymphocytes. Continuous culture of spleen Con A lymphoblasts with chicken IL-15 over two months resulted in an enriched T lymphocyte population expressing the gamma/delta TCR, CD8-alpha, and CD3 cell surface antigens.