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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (20): 140-147.doi: 10.11924/j.issn.1000-6850.2012-1678

所属专题： 生物技术； 水产渔业

• 水产 渔业 • 上一篇    下一篇

斑马鱼G蛋白偶联受体编码基因(DrGPCR)的克隆与表达分析

董超华   

  • 收稿日期:2012-05-02 修回日期:2012-05-09 出版日期:2012-07-15 发布日期:2012-07-15

Cloning and Expression Analysis of a G Protein Coupled Receptor (DrGPCR) From Danio rerioDong Chaohua

  • Received:2012-05-02 Revised:2012-05-09 Online:2012-07-15 Published:2012-07-15

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摘要：

为了探讨硬骨鱼的G蛋白偶联受体的结构和功能。通过RACE技术结合斑马鱼基因组序列分析，鉴定了一个G蛋白偶联受体的编码基因，命名为DrGPCR。该基因全长2251 bp，包括一个1170 bp的开放阅读框，编码389个氨基酸的蛋白。用SMART程序对其氨基酸序列进行分析发现，该蛋白包含有7个跨膜结构域，暗示其为膜蛋白。利用BLAST算法搜索非冗余蛋白数据库发现，DrGPCR蛋白与无脊椎动物GPCR家族的FMRFamide受体以及高等脊椎动物的GPCR139和GPCR142具有高度的氨基酸序列一致性，而且DrGPCR与人GPR139蛋白的三级结构模型具有高度的相似性。序列分析和结构特征都表明DrGPCR是GPCR蛋白家族的一个成员。用邻接法对DrGPCR与绿河豚不同亚族10个GCRP蛋白进行聚类分析发现，DrGPCR属于视紫质受体亚族。通过半定量RT-PCR技术对DrGPCR在斑马鱼各组织的表达进行分析，DrGPCR在脑和肝胰腺中有高水平表达，而在其他被检测的组织没有表达。因此，本研究报道了一个斑马鱼G蛋白偶联受体的编码基因，序列分析和组织表达的结果表明，DrGPCR可能在神经调节和免疫系统中发挥功能。

关键词: 土壤养分, 土壤养分

Abstract:

To explore the structure and function of G protein couple receptors in teleost, a new gene encoding G protein couple receptor were cloned and characterized from zebrafish Danio rerio, and designated as DrGPCR. The full-length cDNA sequence of DrGPCR was 2251 bp containing an 1170 bp of ORF and encoding 389 amino acid residues. SMATR analysis showed that DrGPCR contains seven transmembrane domains, suggesting that it was a membrane protein. BLAST analysis against non-redundant protein database indicated that DrGPCR exhibit high amino acid sequence identity to FMRFamide receptor of invertebrate and GPCR139 or GPCR142 of vertebrates. Moreover, 3D model of DrGPCR was highly similar to that of GPR139 from Homo sapiens. Sequence analysis and structural feature indicated that DrGPCR is a member of GPCR family. Phylogenetic analysis with neighbor joining method indicated that DrGPCR belongs to rhodopsin subgroup. The expression of DrGPCR was examined in various tissues with semi-quantitative RT-PCR. The highest expression of DrGPCR was observed in brain and hepatopancreas. The results all together suggested that DrGPCR might play a role in neuromodulation or immunity.

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