Open Access
How to translate text using browser tools
1 January 1998 SEQUENCE AND CHARACTERIZATION OF PHOCINE INTERLEUKIN 2
Lisi K. M. Shoda, Wendy C. Brown, Allison C. Rice-Ficht
Author Affiliations +
Abstract

To improve assessment of cellular immune responses in seals, northern elephant seal (Mirounga angustirostris) interleukin 2 (IL-2) has been characterized. The gene was cloned and sequenced from a 658 base pair (bp) cDNA generated from total RNA by reverse transcription-polymerase chain reaction (RT-PCR). The sequence encoded a 154 amino acid (aa) polypeptide that included a 20 aa putative signal peptide. Seal IL-2 was found to share considerable identity with published sequences. Nucleotide sequence analysis of phocine (seal) IL-2 with canine, feline, human, trichechine (manatee), bovine and murine sequences demonstrated 93, 92, 86, 82, 78 and 71% identity, respectively. Analysis of the derived amino acid sequences demonstrated 88, 89, 78, 71, 66 and 60% identity, respectively. Interleukin-2 sequence identities appear to reflect evolutionary proximity among the analyzed species, and importantly, those residues identified as critical to IL-2 biological activity and receptor binding are largely conserved. To examine the kinetics of IL-2 mRNA expression, northern elephant seal lymphocytes were stimulated with the mitogen concanavalin A (Con A), and RNA was collected at several time points thereafter. The RT-PCR demonstrated that seal IL-2 mRNA expression peaks in the first 8 hr following Con A stimulation. Lastly, genomic DNA from northern elephant seal, harbour seal (Phoca vitulina) and California sea lion (Zalophus californianus) was used as template to identify and clone genomic IL-2. Partial sequence of the genomic clones demonstrated nearly complete identity among the three species. Sequence identity indicates that probes constructed from the northern elephant seal IL-2 gene will be effective in assessing IL-2 in other pinniped species.

Shoda, Brown, and Rice-Ficht: SEQUENCE AND CHARACTERIZATION OF PHOCINE INTERLEUKIN 2
Lisi K. M. Shoda, Wendy C. Brown, and Allison C. Rice-Ficht "SEQUENCE AND CHARACTERIZATION OF PHOCINE INTERLEUKIN 2," Journal of Wildlife Diseases 34(1), 81-90, (1 January 1998). https://doi.org/10.7589/0090-3558-34.1.81
Received: 12 December 1996; Published: 1 January 1998
KEYWORDS
Cellular immune response
cross-species reactivity
cytokine mRNA expression
interleukin 2
Mirounga angustirostris
northern elephant seal
nucleotide sequence analysis
Back to Top