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家鸡G蛋白偶联受体85基因的克隆与组织表达图谱分析
Molecular Cloning and Tissue Expression of Chicken GPR85 Gene
宋亮,张剑南,刘海坤,莫春横,李娟,王亚军
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作者单位:四川大学生命科学学院,生物资源与生态环境教育部重点实验室
中文关键字:家鸡;GPR85;G蛋白偶联受体;克隆;组织表达
英文关键字:chicken; GPR85; G protein-coupled receptors; clone; tissue expression
中文摘要:G蛋白偶联受体(GPCR)超家族是细胞膜上广泛存在的一类受体,是细胞跨膜信号转导的一类重要受体分子,参与许多生理过程调节。它们中仍有很多至今尚未找到内源性配体,这类受体被称为孤儿型受体。G蛋白偶联受体85(GPR85)是GPCR超家族中孤儿型受体的一员。目前,在非哺乳类脊椎动物中,针对GPR85的研究极少。本研究以家鸡为模型,通过RT-PCR和RACE-PCR等方法从脑中克隆到GPR85基因的cDNA全长序列,揭示其基因结构,并用实时荧光定量PCR (qPCR)的方法探究了该基因在家鸡各组织中的表达情况。结果显示:家鸡GPR85基因位于1号染色体上,由2个外显子组成,其编码区位于第二个外显子上,长为1113bp,可编码1个长为370个氨基酸的七次跨膜受体蛋白。家鸡GPR85与其它脊椎动物(人、小鼠、大鼠、热带爪蟾和斑马鱼)的GPR85具有高度的氨基酸序列一致性(>93%)。qPCR分析发现GPR85基因mRNA在家鸡脑各区域、垂体、肾上腺、精巢中有较高表达水平,而在所检测的其它外周组织中仅有极低的表达水平。本研究首次揭示了家鸡GPR85基因的结构与表达特征,为后续探究GPR85基因在家鸡等非哺乳类中的生理功能奠定基础。
英文摘要:G protein-coupled receptor (GPCR) is a kind of membrane receptor superfamily. It is an important receptor on cell transmembrane to transmit signals, which regulate many important physiological processes. Since the endogenous ligands for some GPCRs have not been identified, thus, these GPCRs have still been classified as orphan receptor. G protein coupled receptor 85 (GPR85) is a member of orphan receptor in GPCR superfamily. Till present, the information regarding on the structure and expression of GPR85 in non-mammalian vertebrates is limited. In this study, using RT-PCR and RACE PCR, we cloned the full cDNA sequence of GPR85 from chicken brain and revealed its gene structure. Furthermore, using quantitative real-time PCR, we also examined the mRNA expression of chicken GPR85 in various chicken tissues. The results showed that chicken GPR85 is located on chromosome 1 and consists of 2 exons. The coding region is located on exon 2. Chicken GPR85 has an open reading frame of 1113bp and encodes a seven-transmembrane receptor with a length of 370 amino acids. Chicken GPR85 shows extremely high amino acid sequence identity (>93%) with that of humans, rats, mice, western clawed frogs and zebrafish. qPCR revealed that chicken GPR85 is highly expressed in the brain regions, anterior pituitary, adrenal glands and testis, and weakly expressed in other remaining tissues examined. To our knowledge, this study represents the first to report the gene structure and expression of GPR85 in chickens and laid the foundation for further exploring the physiological roles of GPR85 in chickens.
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国内统一连续出版物号:51-1193/Q |国际标准出版物号:1000-7083
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