Molecular cloning and functional expression of bottle-nosed dolphin (Tursiops truncatus) interleukin-1 receptor antagonist.
The bottle-nosed dolphin (Tursiops truncatus) interleukin-1 receptor antagonist IL-1ra cDNA was cloned from mitogen-stimulated peripheral blood mononuclear cells (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The sequence of this cDNA showed that dolphin IL-1ra clones contained open reading frames encoding 177 amino acids. Comparison of the deduced amino acids showed that dolphin IL-1ra sequence shared 87.6, 77.9, 77.4, 77.4, 76.4, and 75.8% similarity with the bovine, rabbit, equine, human, mouse, and rat IL-1ra sequences, respectively. Recombinant glutathione S-transferase (GST) dolphin IL-1ra produced in Escherichia coli (E. coli) was purified. This protein suppressed the cytostatic activity of dolphin IL-1beta on A375S2 cells, indicating that the dolphin IL-1ra cDNA obtained in the present study encodes biologically active dolphin IL-1ra.
Inoue
Y
Y
Department of Preventive Veterinary Medicine and Animal Health, Nihon University School of Veterinary Medicine, Nihon University, 1866 Kameino, Fujisawa, 252-8510, Kanagawa, Japan.
Itou
T
T
Jimbo
T
T
Syouji
Y
Y
Ueda
K
K
Sakai
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004289:Dolphins / Q000235:genetics* / Q000276:immunology*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein; D008969:Molecular Sequence Data; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D019105:Random Amplified Polymorphic DNA Technique / Q000662:veterinary; D015375:Receptors, Interleukin-2 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012795:Sialoglycoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology* / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; C507763:IL1RN protein, human; C507764:Il1rn protein, mouse; D053590:Interleukin 1 Receptor Antagonist Protein; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; D012313:RNA