Straubinger A F, Viveiros M M, Straubinger R K
James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Gene. 1999 Aug 20;236(2):273-80. doi: 10.1016/s0378-1119(99)00274-7.
Reverse transcription-polymerase chain reaction (RT-PCR) with interleukin-1alpha (IL-1alpha)-specific primers using total RNA from lipopolysaccharide (LPS)-stimulated lung macrophages resulted in the amplification of two distinct cDNA fragments. Cloning and sequencing of the canine and feline fragments revealed that, except for the absence of a specific 174 nucleotide sequence, the short and the long transcripts were identical. The in-frame 174 nucleotide deletion corresponds to exon 5 of the human and murine IL-1alpha gene, which encodes the cleavage site for calpain, a protein necessary for the processing of the IL-1alpha precursor into mature IL-1alpha. The two transcripts were found in the dog, cat and pig; analysis by RT-PCR, Southern and Northern blot hybridization showed no expression of the shorter IL-1alpha mRNA in equine or bovine macrophages. Expression of the two canine IL-1alpha transcripts was also detected in synovial membranes and was coordinately up-regulated in response to Borrelia burgdorferi infection under both in-vitro and in-vivo conditions.
使用来自脂多糖（LPS）刺激的肺巨噬细胞的总RNA，以白细胞介素-1α（IL-1α）特异性引物进行逆转录聚合酶链反应（RT-PCR），扩增出两个不同的cDNA片段。犬类和猫科动物片段的克隆和测序显示，除了缺少一个特定的174个核苷酸序列外，短转录本和长转录本是相同的。这个读码框内174个核苷酸的缺失对应于人和小鼠IL-1α基因的外显子5，该外显子编码钙蛋白酶的切割位点，钙蛋白酶是将IL-1α前体加工成成熟IL-1α所必需的一种蛋白质。在狗、猫和猪中发现了这两种转录本；通过RT-PCR、Southern和Northern印迹杂交分析表明，马或牛的巨噬细胞中没有较短的IL-1α mRNA表达。在滑膜中也检测到了两种犬类IL-1α转录本的表达，并且在体外和体内条件下，对伯氏疏螺旋体感染的反应中它们都协同上调。
