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家鸡似G蛋白偶联受体119基因的克隆与组织表达分析
Molecular Cloning and Tissue Expression of Chicken GPR119-like Gene
邓秋洋, 张剑南, 王亚军, 李娟*
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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, 成都 610065
中文关键字:家鸡;GPR119b基因;分子克隆;组织表达
英文关键字:chicken; GPR119b gene; molecular cloning; tissue expression
中文摘要:G蛋白偶联受体119(GPR119是A型视紫红质G蛋白偶联受体家族成员。在哺乳动物中,该基因高表达于胰腺β细胞和PP细胞以及小肠组织的内分泌L细胞,参与胰岛素释放调节。本文以家鸡Gallus gallus小肠cDNA为模板,首次克隆到似GPR119新基因,将其命名为cGPR119b基因。结果显示:cGPR119b基因的cDNA全长954 bp,编码具317个氨基酸的前体蛋白。将家鸡GPR119b前体蛋白氨基酸序列与绿头鸭Anas platyrhynchos、绿海龟Chelonia mydas、西部锦龟Chrysemys picta bellii和火鸡Meleagris gallopavo的序列进行比对,分别具有79.9%、61.0%、62.3%、95.5%的相似度。利用反转录PCR和RACE技术,家鸡GPR119b的5'-UTR亦获分离,该片段678 bp。本研究亦采用实时荧光定量PCR探究cGPR119b基因的组织表达,发现其在大脑、肝脏、肾脏、卵巢、精巢、垂体、胰腺、皮肤、脂肪、肺、十二指肠、空肠、回肠、盲肠、直肠等组织中有表达,其中,在肝脏、肾脏、盲肠和精巢的表达量较高,在胰腺、皮肤、脂肪和肺组织中仅有微量表达。本研究为进一步探究cGPR119b基因在家鸡中的生理功能奠定了基础。
英文摘要:G protein-coupled receptor 119 (GPR119) is a member of class A (rhodopsin-type) G protein-coupled receptor family. In mammals, GPR119 gene is found to be highly expressed in pancreatic islet β cells, PP cells and intestine endocrine L cells involving in insulin secretion. In this study, we cloned a chicken GPR119-like gene (named cGPR119b) from intestine and examined its expression in chicken tissues. The results showed that the full-length cDNA of cGPR119b was 954 bp in length and encoding a precursor protein of 317 amino acids. Sequence analysis revealed that chicken GPR119b had high amino acid sequence identity with that of Anas platyrhynchos (79.9%), Chelonia mydas (61.0%), Chrysemys picta bellii (62.3%), and Meleagris gallopavo (95.5%). 5'-RACE PCR assay revealed that the 5'-UTR of cGPR119b is 678 bp. Quantitative real-time PCR assay showed that cGPR119b gene was widely expressed in various chicken tissues including the brain, liver, kidney, intestine, ovary, testis, pituitary, pancreas, skin, fat, lung, duodenum, jejunum, ileum, cecum and rectum. cGPR119b gene was highly expressed in liver, kidneys, cecum and testes, whereas weakly expressed in the pancreas, skin, fat and lung. The present study provided a basis to uncover the physiological roles of cGPR119b gene in chickens.
2018,37(1): 45-50 收稿日期:2017-05-02
DOI:10.11984/j.issn.1000-7083.20170142
分类号:Q959.7;Q78
作者简介:邓秋洋(1990-),硕士,主要从事动物分子遗传学研究,E-mail:13689043376@163.com
*通讯作者:李娟,E-mail:lijuanscuhk@163.com
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