ISOLATION AND CHARACTERIZATION OF A cDNA ENCODING INTERLEUKIN 2 FROM THE FLORIDA MANATEE,TRICHECHUS MANATUS LATIROSTRIS

Abstract

Interleukin‐2 (IL‐2) production is essential for optimal mammalian immune response; therefore, a cDNA encoding IL‐2 was isolated and characterized for the endangered Florida manatee, Trichechus manatus latirostris. The manatee IL‐2 cDNA was cloned by reverse transcription/polymerase chain reaction (PCR), using primers derived from IL‐2 regions conserved among various mammalian species. Manatee IL‐2 cDNA consisted of 450 base pairs encoding a predicted product of 150 amino acid residues and displayed 87%, 82%, 79%, 78%, 75%, and 73% DNA sequence homology and 74%, 69%, 63%, 62%, 61%, and 57% amino acid sequence homology with human, pig, sheep, cow, rat, and mouse IL‐2, respectively.

ISOLATION AND EXPRESSION OF THE INTERLEUKIN‐2 GENE FROM THE KILLER WHALE, ORCINUS ORCA

“…Creation, cloning, and sequencing of killer whale IL-2 specific cDNA-The RNA was converted to single-strand cDNA using the First-Strand cDNA Synthesis Kit manufactured by Clonetech (Palo Alto, CA) following the manufacturer's instructions (details of these procedures can be found in Cashman et al 1996). The cDNA was then used in a 100-pl polymerase chain reaction (PCR), which contained 0.1 volume of the cDNA reaction mixture, 10 p1 1OX PCR buffer (1OX is 100 mM Tris-HCI pH 8.3, 500 mM KCl, 15 mM MgCl,, and 0.01% gelatin), 2 pl each of 10 mM dATP, dCTP, dGTP, and dTTP, 24 pl of a primer mixture containing 600 ng each of the oligonucleotides ATGTACAG-GATGCAACTCCTGT and AGTCAGTGTTGAGATGATGCTTTG.…”

“…In the wild and in captivity these animals must cope with a variety of factors which have the potential to impair their normal immune functions. The isolation and characterization of killer whale IL-2 in this paper and the cloning of manatee (Cashman et al 1996), polar bear (Barber 1996), and dolphin IL-2 (McMahon 1996), manatee (Ness 1995), dolphin (McMahon 1996, and killer whale IL-4 (Ness 1995), and harbor seal, killer whale, and Southern sea otter IL-6 (King et a/. 1996) represent a significant advancement in the establishment of a general knowledge base for marine mammal immunogenetics.…”