鲤鱼白细胞介素-10的克隆、鉴定及表达分析。

Ram Savan, Daisuke Igawa, Masahiro Sakai
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引用次数: 145

摘要

利用鱼头肾IL-10引物,经刀豆蛋白A和脂多糖刺激,从鲤鱼中克隆出白细胞介素-10。cDNA全长1096 bp,包含55 bp的5'非翻译区和498 bp的3'非翻译区。一个543 bp的开放阅读框编码了一个估计有180个氨基酸的蛋白和一个估计有22个氨基酸的信号肽。IL-10的特征基序在鲤鱼序列中是保守的。鲤鱼IL-10的基因组序列为2083 bp,包含5个被4个内含子打断的外显子。除了更紧凑的内含子外,其基因组结构与哺乳动物IL-10相似。经同源性、系统发育和基因组学分析,克隆的鲤鱼基因为IL-10。鲤IL-10在静息期在头、肾、肝、脾和肠中均有表达。在体外脂多糖刺激下,该基因在头肾和肝脏中也有表达。
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Cloning, characterization and expression analysis of interleukin-10 from the common carp, Cyprinus carpio L.

Interleukin (IL)-10 was cloned from the common carp (Cyprinus carpio L.) using IL-10 primers from carp head kidney following stimulation with concanavalin A and lipopolysaccharide. The cDNA consisted of a 1096 bp sequence containing a 55 bp 5' untranslated region and a 498 bp 3' untranslated region. An open reading frame of 543 bp encoded a putative 180 amino acid protein with a putative signal peptide of 22 amino acids. The signature motif of IL-10 is conserved in carp sequence. A 2083 bp genomic sequence of carp IL-10 was found to contain five exons interrupted by four introns. With the exception of much more compact introns, the genomic structure was similar to that of mammalian IL-10. By homology, phylogeny and genomic analyses, the carp gene cloned was designated as IL-10. Carp IL-10 was expressed in head, kidney, liver, spleen and intestine during the resting phase. The gene was also expressed in head kidney and liver following in vitro stimulation with lipopolysaccharide.

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