Yu P W, Chen H T, Czuprynski C J, Schuler L A
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison 53706, USA.
Cytokine. 1997 Jan;9(1):1-8. doi: 10.1006/cyto.1996.0129.
The purpose of this study was to clone and characterize the bovine IL-1rII. Using a human IL-1rII probe, three putative exons of the bovine IL-1rII were identified from a bovine genomic library. The full length cDNA for bovine IL-1rII was cloned from a bovine endometrial cDNA library using oligonucleotides designed from those exons. The nucleotide sequence of the bovine IL-1rII cDNA has 79, 69 and 69% identity with those of human, mouse, and rat IL-1rII, respectively. The bovine IL-1rII cDNA contains an open reading frame encoding 400 amino acids with a predicted amino acid sequence of 71, 58, and 59% identity with those of human, mouse and rat IL-1rII, respectively. Transient transfection of COS cells with the cloned bovine IL-1rII cDNA resulted in specific binding of bovine IL-1 beta, with a Kd of 1.82 x 10(-10) M. At least two transcripts of bovine IL-1rII were identified from bovine peripheral blood mononuclear cells (PBMCs), liver, and fetal thymocytes by Northern blot analysis. The relative proportion of these transcripts varied with cell type. In addition, bovine IL-1rII transcripts were detected by RT-PCR in stimulated and unstimulated neutrophils, fibroblasts, and splenocytes. The results of this study provide the first characterization of bovine IL-1rII at a molecular level.
本研究的目的是克隆并鉴定牛白细胞介素-1受体II型（IL-1rII）。使用人IL-1rII探针，从牛基因组文库中鉴定出牛IL-1rII的三个推定外显子。利用从这些外显子设计的寡核苷酸，从牛子宫内膜cDNA文库中克隆出牛IL-1rII的全长cDNA。牛IL-1rII cDNA的核苷酸序列与人类、小鼠和大鼠的IL-1rII分别具有79%、69%和69%的同源性。牛IL-1rII cDNA包含一个编码400个氨基酸的开放阅读框，其预测的氨基酸序列与人类、小鼠和大鼠的IL-1rII分别具有71%、58%和59%的同源性。用克隆的牛IL-1rII cDNA瞬时转染COS细胞，导致牛白细胞介素-1β（IL-1β）的特异性结合，解离常数（Kd）为1.82×10⁻¹⁰M。通过Northern印迹分析，从牛外周血单核细胞（PBMC）、肝脏和胎儿胸腺细胞中鉴定出至少两种牛IL-1rII转录本。这些转录本的相对比例因细胞类型而异。此外，通过逆转录聚合酶链反应（RT-PCR）在受刺激和未受刺激的中性粒细胞、成纤维细胞和脾细胞中检测到牛IL-1rII转录本。本研究结果首次在分子水平上对牛IL-1rII进行了鉴定。
