牛白细胞介素-1受体拮抗剂cDNA的酶促扩增与表达
Enzymatic amplification and expression of bovine interleukin-1 receptor antagonist cDNA.
作者信息
Kirisawa R, Fukuda T, Yamanaka H, Hagiwara K, Goto M, Obata Y, Yoshino T, Iwai H
机构信息
Department of Veterinary Microbiology, School of Veterinary Medicine, Rakuno Gakuen University, Hokkaido, Japan.
出版信息
Vet Immunol Immunopathol. 1998 Apr 16;62(3):197-208. doi: 10.1016/s0165-2427(98)00099-3.
cDNA generated from lipopolysaccharide-stimulated bovine peripheral blood mononuclear cells was used to amplify and clone the bovine interleukin-1 receptor antagonist (IL-1ra) using primers derived from semi-conserved regions between human and mouse IL-1ra sequences. 5' and 3' terminal sequences of bovine IL-1ra were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of bovine IL-1ra demonstrated 80%, 78%, 78%, 77% and 76% homology with human, mouse, rat, rabbit and equine sequences, respectively. Recombinant bovine IL-1ra produced in Escherichia coli suppressed the growth inhibitory activity of bovine IL-1beta on A375 cells in a dose-dependent manner, indicating that the present bovine IL-1ra cDNA encodes biologically active proteins.
从脂多糖刺激的牛外周血单核细胞生成的互补DNA（cDNA），使用源自人和小鼠白细胞介素-1受体拮抗剂（IL-1ra）序列之间半保守区域的引物，用于扩增和克隆牛IL-1ra。通过cDNA末端的5'和3'快速扩增来扩增牛IL-1ra的5'和3'末端序列。推导的牛IL-1ra氨基酸序列分别与人、小鼠、大鼠、兔和马的序列具有80%、78%、78%、77%和76%的同源性。在大肠杆菌中产生的重组牛IL-1ra以剂量依赖性方式抑制牛IL-1β对A375细胞的生长抑制活性，表明目前的牛IL-1ra cDNA编码具有生物活性的蛋白质。
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