京海黄鸡G蛋白偶联受体1基因生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2017.08.001

摘要/Abstract

摘要：

试验旨在研究参与激素调节并调控卵巢功能的可能基因，挑选候选基因G蛋白偶联受体1（G protein-coupled receptor 1），深入研究其是否参与激素调节。根据转录组测序获得的京海黄鸡GPR1基因的CDS序列设计一对引物，利用实时荧光定量RT-PCR方法，检测GPR1基因在高、低产京海黄鸡卵巢组织中的表达情况，并与生物信息学进行关联，使用多种生物软件和在线工具对目的序列进行生物信息学分析，分析GPR1基因与其他物种的同源性、蛋白质的理化性质、蛋白质序列的跨膜区、亚细胞定位、亲水性、潜在的磷酸化位点、保守结构域及该基因所编码蛋白质的二、三级结构。结果显示，京海黄鸡与GenBank中原鸡、人、牛、野猪、黑猩猩、家兔、马、绵羊、藏羚羊、大鼠、小鼠、斑马鱼的同源性分别为100.0%、69.0%、69.1%、69.3%、68.9%、69.6%、69.2%、68.8%、68.9%、67.5%、69.0%和50.3%；氨基酸分析发现GPR1蛋白为疏水性蛋白，分子质量为40.41 ku，理论等电点为6.91，为7次跨膜蛋白；亚细胞定位显示，该蛋白属于非分泌蛋白；预测该蛋白含有25个潜在的磷酸化位点，1糖基化位点；二级结构以无规则卷曲为主，三级结构为7次跨膜螺旋结构；组织表达分析显示，GPR1基因在低产组的表达情况极显著高于高产组（P<0.01）。本试验结果将有利于GPR1基因功能的进一步分析，为研究该基因对京海黄鸡产蛋性状的调控机理提供理论依据。

关键词: 京海黄鸡; G蛋白偶联受体1; 生物信息学分析; 蛋白; 组织表达

Abstract:

This study was aimed to investigate the possible genes that involved in hormone regulation and regulate ovarian function, and selected G protein-coupled receptor 1 to in-depth study whether it was involved in hormone regulation.In this study, a pair of primers was designed based on the CDS sequence of GPR1 gene of Jinghai Yellow chicken which according to transcriptome sequencing.Real-time RT-PCR was used to detect the expression of GPR1 gene in the ovary of high and low-production Jinghai Yellow chicken. Bioinformatics analysis of GPR1 gene was performed by using combined biological softwares and online tools. The homology of GPR1 gene with other species, the physicochemical properties of protein, the transmembrane region, subcellular localization, hydrophilicity, potential phosphorylation site, conserved domain of protein sequence, secondary structure and tertiary structure of GPR1 protein were analyzed. The results showed that GPR1 gene of Jinghai Yellow chicken shared 100.0%, 69.0%, 69.1%, 69.3%, 68.9%, 69.6%, 69.2%, 68.8%, 68.9%, 67.5%, 69.0% and 50.3% identity with Gallus gallus, Homo sapiens, Bos taurus,Sus scrofa,Pan troglodytes,Leporidae,Equus caballus,Ovis aries, Pantholops hodgsonii, Rattus norvegicus,Mus musculus and Danio rerio, respectively.GPR1 protein structure showed that the molecular weight was 40.41 ku and pI was 6.91, including seven transmembrane segments which were consistent with the transmembrane analysis. Subcellular localization showed that GPR1 protein belonged to non-secretory protein. The predicted results showed that the protein contained 25 potential phosphorylation sites and 1 glycosylation sites. The secondary structure of GPR1 protein was mainly composed of random coil. The tertiary structure of domain area of GPR1 protein showed a transmembrane helix and single strand structure. The result of tissue expression showed that the expression of GPR1 gene in the low-production group was extremely significantly higher than the high-production group (P<0.01).The results of the study would be beneficial to the further analysis of the function of GPR1 gene, and provided a theoretical basis for the further study of the gene on the regulation mechanism of laying traits of Jinghai Yellow chicken.

Key words: Jinghai Yellow chicken; G protein-coupled receptor 1; bioinformatics analysis; protein; tissue expression

中图分类号:

S831

袁曼曼, 张涛, 张向前, 李婷婷, 凌娇娇, 吴鹏飞, 王金玉, 施会强. 京海黄鸡G蛋白偶联受体1基因生物信息学及组织表达分析[J]. 《中国畜牧兽医》, 2017, 44(8): 2215-2225.

YUAN Man-man, ZHANG Tao, ZHANG Xiang-qian, LI Ting-ting, LING Jiao-jiao, WU Peng-fei, WANG Jin-yu, SHI Hui-qiang. Bioinformatics and Tissue Expression Analysis of G Protein-coupled Receptor 1 Gene in Jinghai Yellow Chicken[J]. , 2017, 44(8): 2215-2225.

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