| 斑马鱼BLT1-like基因的克隆和表达分析 |
| |
| 引用本文： | 唐宇飞,刘晓红,常馨月,吴海珍,张元兴.斑马鱼BLT1-like基因的克隆和表达分析[J].水产学报,2015,39(12):1753-1762. |
| |
| 作者姓名： | 唐宇飞 刘晓红 常馨月 吴海珍 张元兴 |
| |
| 作者单位： | 华东理工大学 生物工程学院,华东理工大学 生物工程学院；上海海洋动物疫苗工程技术研究中心,华东理工大学 生物工程学院,华东理工大学 生物工程学院,华东理工大学 生物工程学院；上海海洋动物疫苗工程技术研究中心 |
| |
| 基金项目： | 国家自然科学基金(31372550)，国家鲆鲽类产业技术体系(CARS-50-G07) |
| |
| 摘 要： | 利用RACE技术克隆得到了斑马鱼(Danio rerio)白三烯B4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,其编码的两个蛋白均属于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。进一步将两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基因在胚胎发育12 h显著上调,mRNA表达量上升至1 h的18倍,而BLT1-like2基因在胚胎发育24 h发生显著上调,表达量上升至1 h的34倍；而在斑马鱼成鱼中两个基因在心脏和肝脏中表达量相对较高,而肠、皮和眼睛中表达量相对较低。本研究的结果说明斑马鱼中可能存在多个BLT1基因,并为鱼类BLT1基因的克隆和功能鉴定提供基础。
|
| 关 键 词： | 斑马鱼 BLT1 基因克隆 基因表达 |
| 收稿时间： | 2015/6/11 0:00:00 |
| 修稿时间： | 9/1/2015 12:00:00 AM |
Cloning and expression analysis of BLT1-like genes from zebrafish(Danio rerio) |
| |
| TANG Yufei,LIU Xiaohong,CHANG Xinyue,WU Haizhen and ZHANG Yuanxing.Cloning and expression analysis of BLT1-like genes from zebrafish(Danio rerio)[J].Journal of Fisheries of China,2015,39(12):1753-1762. |
| |
| Authors: | TANG Yufei LIU Xiaohong CHANG Xinyue WU Haizhen and ZHANG Yuanxing |
| |
| Institution: | State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China,State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;Shanghai Collaborative Innovation Center for Biomanufacturing Technology, Shanghai 201424, China,State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China,State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;Shanghai Collaborative Innovation Center for Biomanufacturing Technology, Shanghai 201424, China |
| |
| Abstract: | Using RACE (Rapid Amplification of cDNA Ends) technology, two leukotriene B4 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), occupy 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 well membrane-bound property was found. And western blotting experiment verified their specific interactions with human BLT1 polyclonal antibodies. Besides, the results of RT-qPCR 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 low expression level in the intestine, skin and eye. Conclusively, our study indicates that there might exit more than one BLT1 gene in zebrafish and this will be helpful to clone and characterize BLT1 genes from other fishes. |
| |
| Keywords: | zebrafish BLT1 gene cloning gene expression |
|
| 点击此处可从《水产学报》浏览原始摘要信息 |
| 点击此处可从《水产学报》下载免费的PDF全文 |
|
