Shoji Y, Inoue Y, Sugisawa H, Itou T, Endo T, Sakai T
Department of Preventive Veterinary Medicine and Animal Health, Nihon University School of Veterinary Medicine, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan.
Vet Immunol Immunopathol. 2001 Oct;82(3-4):183-92. doi: 10.1016/s0165-2427(01)00353-1.
Bottlenose dolphin tumor necrosis factor alpha (doTNF-alpha) cDNA was cloned by reverse transcription polymerase chain reaction (RT-PCR) and the nucleic and deduced amino acid sequences were determined. The sequence of the cDNA clones shows that doTNF-alpha has an open reading frame of 699bp encoding 233 amino acids. The nucleic acid sequence of doTNF-alpha indicates 90, 88, 87, and 79% similarity with the cattle, pig, human, and mouse TNF-alpha gene, respectively. Based on the analysis of human and mouse TNF-alpha molecules, doTNF-alpha is processed to a mature protein with 157 amino acids. The 233 amino acids precursor has a hydrophobic region that could serve as a transmembrane domain. The recombinant doTNF-alpha expressed in Escherichia coli as a glutathione S-transferase fusion protein reacted with anti-human TNF-alpha antibody and exerted cytotoxity to the TNF-alpha sensitive murine cell line L929.
