扬州鹅NPFFR1基因CDS区克隆、生物信息学分析及组织表达检测  

Cloning, bioinformatics and tissue expression analysis of NPFFR1 gene coding sequence region in Yangzhou goose

作　　者：汪琴 Melak Sherif 韦伟[1] 张立凡[1] 陈杰[1] WANG Qin;Melak Sherif;WEI Wei;ZHANG Lifan;CHEN Jie(College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物科技学院

出　　处：《畜牧与兽医》2019年第11期1-10,共10页Animal Husbandry & Veterinary Medicine

基　　金：江苏省农业重大新品种创制项目(PZCZ201738)

摘　　要：本研究旨在对鹅促性腺激素抑制激素受体基因(NPFFR1)进行克隆及生物信息学分析,并检测其在鹅不同组织中的表达。利用鸡NPFFR1基因为种子序列,采用RT-PCR方法克隆出扬州鹅NPRRF1基因,并通过相关软件对基因序列进行生物信息学分析。结果显示:共发现扬州白鹅NPFFR1基因的两种剪接形式:剪接体1的ORF大小为1 200 bp,编码399个氨基酸;剪接体2的ORF大小为432 bp,编码143个氨基酸,相对剪接体1缺失31 bp。对其进行生物信息学分析发现,鹅NPFFR1基因与鸡NPFFR1基因CDS序列同源性为92.8%,其产物是一种不稳定、疏水性蛋白,主要分布在细胞膜上;蛋白质跨膜结构预测显示鹅NPFFR1蛋白含有7个跨膜结构,属于跨膜蛋白,无信号肽片段。对产蛋末期母鹅10个组织样品的cDNA实时定量PCR结果显示,NPFFR1基因在各组织中均有表达,在下丘脑中表达最高,其次是腹脂、小肠、卵巢、胃、垂体、心脏、肌肉,在肾脏以及肝脏中表达很低。研究结果为今后探究鹅NPFFR1蛋白的相关功能及与其他蛋白质的相互作用提供了理论依据,同时为扬州鹅生殖轴调控产蛋性能的机理研究提供参考。This study aims to clone NPFFR1 gene of Yangzhou goose, analyze the sequence characteristic by bioinformatics software, and detect the expression of NPFFR1 gene in different tissues of Yangzhou goose. Using the NPFFR1 gene of Gallus(Gen Bank: NM204362.1) as the seed sequence, the complete CDS of goose NPFFR1 gene was successfully cloned by RT-PCR, and then network resources and relevant softwares were used to analyze the sequence characteristic of the cloned gene. The results showed that there were two splicing forms of NPFFR1 gene in Yangzhou goose. The open reading frame size of splicing 1 was 1200 bp, encode 399 amino acids;the cDNA length of splicing 2(31 bP was missing from splice 1) was 1169 bp, and its ORF was 432 bp in length and encodes 143 amino acids. Bioinformatics analysis revealed that the homology of this NPFFR1 gene and chicken NPFFR1 gene CDS sequence was 92.8%. Its product was an unstable and hydrophobic protein, mainly distributed on the cell membrane, and the prediction result showed that the goose NPFRF1 protein is a transmembrane protein containing seven transmembrane structures without signal peptide fragments. Real-time quantitative PCR was performed using the cDNA samples of ten goose tissues at the late period of laying. The results showed that the goose NPFFR1 gene was expressed in all tissues and highest in the hypothalamus, followed by abdominal fat, small intestine, ovary, stomach and pituitary, heart, muscle, and the expression in the kidneys and liver is very low. The results contribute to the exploration of NPFFR1 protein properties and its interaction with other proteins, and provide information for the future research on the regulation mechanism of Yangzhou goose reproductive axis.

关 键 词：扬州鹅 NPFFR1基因 克隆 生物信息学分析 

分 类 号：S835[农业科学—畜牧学]