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[目的]进行鲤鱼白细胞介素-1β(IL-1β)全长cDNA的克隆、鉴定及其差异表达分析。[方法]利用DD-RTPCR方法获得差异表达cDNA片段,对有丝分裂原刺激的鲤鱼外周血白细胞cDNA文库进行筛选,克隆了鲤鱼IL-1β的全长cDNA,并进行了序列分析和差异表达分析。[结果]获得的阳性克隆含有1个大小为831 bp编码276个氨基酸的完整开放阅读框。聚类分析表明,鲤鱼IL-1β氨基酸序列与日本鲤鱼紧密聚为一支,氨基酸序列的同源性达95%,之后聚类顺序依次为鲫鱼、斑马鱼、猪、牛、马、人和小鼠。差异表达分析表明,经有丝分裂原刺激后前期(4 h)白细胞中IL-1β的表达量显著增大,但随着时间推移(12,24 h)并非一直较同期大,表达量总体趋势成峰形。[结论]为进一步研究IL-1β在体内的表达方式、功能特点和调控机理以及在炎症反应、应急反应和免疫应答的作用机制奠定了基础。
[Objective] The research aimed to clone, identify and differentiate the full-length cDNA of IL-1β in common carp. [Method] The differentially expressed cDNA fragments were obtained by DD-RTPCR method. The mitochondria-stimulated carp peripheral blood leucocyte cDNA library was screened. The full-length cDNA of IL-1β in common carp was cloned and sequenced and differentially expressed. [Result] The positive clone obtained contained a complete open reading frame with a size of 831 bp encoding 276 amino acids. The results of cluster analysis showed that the amino acid sequence of IL-1β was closely related to Japanese carp, and the homology of the amino acid sequence was 95%. The order of clustering was carp, zebra fish, pig, cow, horse, human and small mouse. Differential expression analysis showed that the expression of IL-1β in leukocytes stimulated by mitogen was significantly increased, but it was not always higher than that of the same period over time (12 and 24 h), and the overall trend of expression peaked shape. [Conclusion] This study lays the foundation for further study on the expression, function and regulation of IL-1β in vivo and the mechanism of action in response to inflammation, emergency and immune response.