Molecular characterization and expression of caprine (Capra hircus) interleukin-18 cDNA.
Interleukin 18 (IL-18) has been identified as a potent upstream cytokine required for upregulation of IFN-gamma secretion that plays a crucial role in polarization of Th1 type of immune response. Considering the potential applications of the cytokine in immunomodulation, it has been characterized in many livestock species including cattle, equines, canines, felines and porcines. In this paper we report the isolation, cloning sequencing and expression of caprine precursor IL-18. Full-length caprine IL-18 cDNA was isolated from mitogen-stimulated adherent peripheral blood mononuclear cells using reverse transcription polymerase chain reaction (RT-PCR). The cDNA contained an open reading frame of 579 bp encoding a putative polypeptide of 192 amino acids. Deduced amino acid sequence of caprine IL-18 showed varying amino acid identity with the published sequences of other domestic ruminant species ranging from 94.3% to 96.9%, while it shared over 78% aa identity with other domestic animals. Pairwise multiple aligned sequences showed a deletion of Glu31in caprine IL-18 unlike in other species. Recombinant caprine IL-18 was produced in Escherichia coli, which cross-reacted with two antiporcine IL-18 monoclonal antibodies.
Hosamani
M
M
Indian Veterinary Research Institute, Mukteswar-263 138, National Uttaranchal State, India.
Mondal
B
B
Muneta
Y
Y
Rasool
T J
TJ
D000818:Animals; D000829:Animals, Domestic / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression / Q000235:genetics*; D006041:Goats / Q000235:genetics* / Q000276:immunology; D020382:Interleukin-18 / Q000235:genetics* / Q000276:immunology; D016366:Open Reading Frames / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D018076:DNA, Complementary; D020382:Interleukin-18