Inoue Y, Itou T, Jimbo T, Syouji Y, Ueda K, Sakai T
Department of Preventive Veterinary Medicine and Animal Health, Nihon University School of Veterinary Medicine, Nihon University, 1866 Kameino, Fujisawa, 252-8510, Kanagawa, Japan.
Vet Immunol Immunopathol. 2001 Jan 26;78(2):131-41. doi: 10.1016/s0165-2427(00)00263-4.
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.
利用逆转录聚合酶链反应（RT-PCR），从有丝分裂原刺激的外周血单核细胞（PBMC）RNA中克隆出瓶鼻海豚（宽吻海豚）白细胞介素-1受体拮抗剂IL-1ra的cDNA。该cDNA序列表明，海豚IL-1ra克隆包含编码177个氨基酸的开放阅读框。对推导氨基酸的比较显示，海豚IL-1ra序列与牛、兔、马、人、小鼠和大鼠的IL-1ra序列的相似性分别为87.6%、77.9%、77.4%、77.4%、76.4%和75.8%。对在大肠杆菌（E. coli）中产生的重组谷胱甘肽S-转移酶（GST）海豚IL-1ra进行了纯化。该蛋白抑制了海豚IL-1β对A375S2细胞的细胞生长抑制活性，表明本研究中获得的海豚IL-1ra cDNA编码具有生物活性的海豚IL-1ra。
