豫西脂尾羊INSIG1基因克隆、序列分析及组织表达  

Cloning,Sequence Analysis and Tissue Expression of INSIG1 Gene in Yuxi Fat-tailed Sheep

作　　者：权凯 宋飞虎 李君 QUAN Kai;SONG Feihu;LI Jun(Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China)

机构地区：[1]河南牧业经济学院动物科技学院,郑州450046

出　　处：《中国畜牧兽医》2020年第6期1781-1790,共10页China Animal Husbandry & Veterinary Medicine

基　　金：河南省科技攻关专项(192102110175);国家自然科学基金项目(31601915)。

摘　　要：胰岛素诱导基因1(INSIG1)是脂质合成与分解的重要调控基因,为了研究豫西脂尾羊INSIG1基因序列特征及其组织表达规律,试验采用Trizol法提取组织样RNA,RT-PCR扩增后克隆得到INSIG1基因序列并进行分析;采用实时荧光定量PCR法检测INSIG1 mRNA表达情况,并对结果进行比较分析。试验成功克隆了豫西脂尾羊INSIG1基因,其编码区长831 bp,编码276个氨基酸;基因的同源性分析表明,豫西脂尾羊INSIG1基因编码区与绵羊(XM_015095466.2)的亲缘关系相似性达99.64%,编码氨基酸序列的相似性达99.28%;蛋白理化性质分析表明,其分子质量为29.58 ku,理论等电点(pI)为9.07,属于稳定的碱性疏水性蛋白;跨膜结构、信号肽和亚细胞定位分析表明,该蛋白包含5个跨膜结构,没有信号肽,主要分布在细胞质;蛋白质三级结构预测发现,INSIG1蛋白结构含有6个α-螺旋和部分无规则卷曲;实时荧光定量PCR结果表明,INSIG1基因在肝脏中表达量最高,其次为肺脏、小肠和尾脂,肌肉中表达量最低。本研究完善了豫西脂尾羊的数据库,为INSIG1基因的功能及其在肉羊脂肪沉积过程中的作用机制提供了依据。Insulin induced gene 1(INSIG1)was an important regulatory gene of lipid synthesis and lipolysis.This study was aimed to clone INSIG1 gene,analyze the sequences by bioinformatics software,and measure the expression of INSIG1 gene in different tissues of Yuxi Fat-tailed sheep.The tissue RNA was extracted by Trizol method,and the INSIG1 gene sequence was obtained by RT-PCR and cloning.The expression of INSIG1 mRNA was detected by Real-time PCR.The results showed that the complete CDS of sheep INSIG1 gene was successfully cloned which was 831 bp and encoded 276 amino acids.The homology analysis showed that the INSIG1 coding region in Yuxi Fat-tailed sheep was 99.64%similar to that of sheep(XM_015095466.2),and the similarity of coding amino acid sequence was 99.28%.The physical and chemical properties of INSIG1 protein showed that the molecular weight was 29.58 ku and the theoretical isoelectric point(pI)was 9.07,which belonged to stable alkaline hydrophobic protein.There was 5 transmembrane structures and no signal peptide in INSIG1 protein,mainly located in cytoplasm.The tertiary structure of INSIG1 protein mainly consisted of 6 alpha helices and random coils.Real-time PCR results showed that the expression of INSIG1 gene in liver was obviously higher than other tissues,which followed by lung,intestine,and adipose of tail,and the lowest was in muscle.This study improved the database of Yuxi Fat-tailed sheep,and provided basic data for further study on the function of INSIG1 gene in fat deposition and regulation of mutton quality in sheep.

关 键 词：豫西脂尾羊 INSIG1基因 克隆 组织表达 

分 类 号：S826[农业科学—畜牧学]