齐口裂腹鱼白介素-1β基因的克隆及表达谱

Cloning and Expression Profile of Interleukin-1β Gene in Schizothorax prenanti

探讨齐口裂腹鱼白介素-1β(Interleukin-1β,IL-1β)基因的基本特点。用RT-PCR方法以齐口裂腹鱼脾脏c DNA作为模板扩增IL-1β基因,用生物信息学软件分析其序列,用RT-PCR方法检测IL-1β基因在齐口裂腹鱼6种组织的表达情况。克隆获得的IL-1β序列长1 252 bp,共编码276个氨基酸。Gen Bank序列号为KU886235。IL-1β蛋白相对分子量为31.25 k D,等电点5.45,预测为亲水性蛋白。二级结构主要结构元件为随机卷曲和延伸链,有4个N-糖基化位点和16个磷酸化位点。NJ法系统进化树显示,齐口裂腹鱼与鲤鱼、鲫鱼的亲缘关系最近。组织表达分析显示,IL-1β基因主要在脾脏和肾脏中表达。这为深入研究鱼IL-1β的生物学功能提供理论依据。

This work aims to characterize interleukin-1β（ IL-1β） gene of Schizothorax prenanti. First, RT-PCR was used to amplify IL-lJ] gene with cDNA of S. prenanti spleen as template. Then, the IL-1β gene sequence was analyzed with various bioinformatics tools, and the expressions in 6 tissues of S. prenanti were detected used RT-PCR. As results, the length of IL-1β gene was 1 252 bp, and it encoded 276 amino acids. Its GenBank accession number was KU886235, the molecular weight of IL-1β was 31.25 kD, and pI was 5.45. It was induced as a hydrophilic protein, and its secondary structure mainly consisted of random coil and extended strand. There were 4 N-glycosylation sites and 16 phosphorylation sites in this protein. Phylogenetic relationships revealed that S. prenanti presented a closest genetic relationship with Cyprinus carpio and Carassius auratus. The analysis of tissue expression indicated that IL-1β gene was mainly expressed in the spleen and kidney. It provides theoretical evidence for further studying the biological function of IL-1β gene.