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[目的]获取鲤鱼全长IL-10(interleukin 10)cDNA,并对其序列进行分析。[方法]利用DD-RTPCR(differential display RT-PCR)方法获得差异表达IL-10 cDNA片段,以地高辛标记做为探针,对有丝分裂原刺激的鲤鱼外周血白细胞cDNA文库进行核酸杂交筛选,克隆鲤鱼IL-10全长cDNA,并对该序列进行序列分析和同源性比较。[结果]鲤鱼全长IL-10 cDNA共1 117 bp,包含55 bp的5’端非编码区,一个540 bp的编码179个氨基酸的开放阅读框及522 bp的3’端非编码区,其中,3’非编码区包含3个mRNA不稳定基序“ATTTA”;该蛋白序列具有IL-10家族的典型序列特征;序列同源性比较表明,所获得的序列与GenBank上登录的鲤鱼IL-10基因同源性为89.1%。[结论]该试验为进一步研究IL-10在体内的表达方式、功能特点、调控机理及其在炎症反应和免疫应答中的作用机制奠定了基础。
[Objective] The research aimed to obtain the full length IL-10 (interleukin 10) cDNA of common carp and analyze its sequence. [Method] DD-RTPCR (differential display RT-PCR) method was used to obtain differentially expressed cDNA fragment of IL-10. Digoxigenin was used as a probe to screen the cDNA library of mitogens stimulated carp peripheral white blood cells. The full length cDNA of common carp IL-10 was cloned and sequenced and compared for homology. [Result] The total length of IL-10 cDNA in common carp was 1 117 bp in length, including a 55 bp 5 ’untranslated region, a 540 bp open reading frame encoding 179 amino acids and a 3’ non-coding region of 522 bp , 3 ’non-coding region contains three mRNA instability motifs’ ATTTA’; This protein sequence has the typical sequence features of the IL-10 family; Sequence homology comparison shows that the obtained sequence with the carp registered GenBank IL-10 gene homology was 89.1%. [Conclusion] This study lays the foundation for further study of IL-10 expression pattern, functional characteristics, regulatory mechanism and its mechanism of action in inflammatory response and immune response in vivo.