中文摘要：

为了研究白三烯B_4受体在斑马鱼发育和免疫中的功能,本实验利用RACE技术克隆得到了斑马鱼白三烯B_4受体样（BLT1-like）基因BLT1-like1和BLT1-like2的全长cDNA序列,其分别编码339和356个氨基酸。序列和结构分析发现,其编码的2个蛋白均属于G蛋白偶联受体家族视紫红质蛋白亚族,并具有典型的G蛋白偶联受体的特征,与人的BLT1同源度达31%以上。进一步将2个基因与绿色荧光蛋白融合表达,发现其具有较好的细胞膜定位功能;Western印迹检测也证实,重组表达细胞的全细胞蛋白可与人源BLT1的多克隆抗体发生特异性相互作用。此外,荧光定量PCR分析结果显示,在斑马鱼幼鱼发育过程中,BLT1-like1在胚胎发育12 h显著上调,mRNA表达量上升至1 h时的18倍,而BLT1-like2在胚胎发育24 h发生显著上调,表达量上升至1 h时的34倍;斑马鱼成鱼中的2个基因在心脏和肝脏中表达量相对较高,而肠道、皮肤和眼睛中表达量相对较低。说明斑马鱼体内可能存在多个BLT1基因,实验结果为鱼类BLT1基因的克隆和功能鉴定提供基础。

英文摘要：

In order to research the functions of BLT1 in the development and vaccination process of zebrafish,two leukotriene B_4 receptor 1-like（ BLT1-like） genes,BLT1-like1 and BLT1-like2,from zebrafish were cloned,which encoded 339 and 356 amino acids,respectively. Through sequence and structure analysis,it was found that both of the two proteins belong to rhodopsin subfamily of G protein coupled receptors（ GPCR）,with typical features of GPCR and have a moderate identity of above 31% with human BLT1.Additionally,when fused with enhanced green fluorescent protein（ EGFP） and expressed in CHO cells,a good membrane-bound property was found. And Western blotting experiment verified their specific interactions with human BLT1 polyclonal antibodies. Besides,the results of RT-q PCR showed that during the embryo development of zebrafish,the expression of BLT1-like1 was up-regulated significantly at 12 h,which was 18 times higher than that of 1 h. While the expression level of BLT1-like2 changed sharply at 24 h,which was 34 times higher than that of 1 h. In addition,both of the two genes had a relatively high expression level in the heart and liver,while with a relatively lowexpression level in the intestine,skin and eye. Conclusively,our study indicates that there might exist more than one BLT1 gene in zebrafish and this will be helpful to clone and characterize BLT1 genes from other fishes.