湖羊SREBP1基因编码区序列克隆及表达分析  

Sequence cloning and expression analysis of SREBP1 gene in Hu sheep

作　　者：李樊 舒嘉傲 李隐侠[2] 孟春花[2] 张晨俭 张俊[2] 钱勇[2] 曹少先[1,2] LI Fan;SHU Jia-ao;LI Yin-xia;MENG Chun-hua;ZHANG Chen-jian;ZHANG Jun;QIAN Yong;CAO Shao-xian(College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China;Institute of Animal Science,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]南京农业大学动物科技学院,江苏南京210095 [2]江苏省农业科学院畜牧研究所,江苏南京210014

出　　处：《江苏农业学报》2022年第3期721-729,共9页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省农业科技自主创新基金项目[CX(20)3013];国家自然科学基金项目(31902155)。

摘　　要：SREBP1是一个重要的核转录因子,参与调控胆固醇和脂肪酸等合成和代谢,本研究旨在基于获得湖羊SREBP1基因序列并分析其组织表达谱和卵巢中的表达定位,了解其在湖羊中可能的生物学功能。本研究利用RT-PCR技术对周岁湖羊卵巢组织中SREBP1基因编码区序列进行克隆和测序,用生物信息学软件对序列进行拼接、同源比对和功能预测分析,荧光定量PCR检测其在湖羊组织中的表达模式,免疫组织化学染色法鉴定其在卵巢组织中的定位。结果显示:SREBP1基因在湖羊卵巢组织中有2种剪接体,对应编码区长度分别为3369 bp和3441 bp,分别编码1122个和1146个氨基酸残基;SREBP1在哺乳动物中相对保守,含有经典的bHLH-Zip结构域,通过识别E-boxes或者固醇调节元件调控靶基因的表达;组织表达谱分析结果显示,SREBP1在湖羊组织中广泛表达,提示SREBP1基因可能参与调控湖羊多种生物学过程。免疫组织化学染色法分析结果显示,SREBP1基因在湖羊卵巢腔前卵泡和有腔卵泡中均有表达,主要定位于颗粒细胞、卵泡膜细胞和黄体细胞中,推测SREBP1可能在湖羊繁殖调控中发挥重要的作用。这些结果为进一步研究湖羊SREBP1基因的功能提供了基础。SREBP1 is an important nuclear transcription factor,which is involved in regulating the synthesis and metabolism of cholesterol and fatty acids and other biological functions.The purpose of this study is to understand the possible biological functions of SREBP1 gene in Hu sheep based on its coding sequence,tissue expression profile and location in ovary.In this study,the coding region of SREBP1 gene in Hu sheep ovarian tissue was cloned and sequenced by RT-PCR.Bioinformatics software was used for splicing,homologous alignment and functional prediction analysis.The expression pattern in Hu sheep tissue was detected by real-time PCR,and its localization in ovarian tissue was identified by immunohistochemistry.The results showed that SREBP1 gene had two kinds of spliceosomes in Hu sheep ovary,and the length of corresponding coding regions was 3369 bp and 3441 bp,and encoding 1122 and 1146 amino acid residues,respectively.SREBP1 was relatively conserved in mammals and contained the classical bHLH-Zip domain.In addition it could regulate the expression of target genes by identifying E-boxes or sterol regulatory elements.The results of tissue expression profile analysis showed that SREBP1 was widely expressed in Hu sheep,suggesting that SREBP1 gene may be involved in regulating a variety of biological processes in Hu sheep.The results of immunohistochemical staining showed that SREBP1 gene was expressed in both antral follicles and preantral follicles of Hu sheep ovary,mainly located in granulosa cells,theca cells and luteal cells,suggesting that SREBP1 may play an important role in the reproductive regulation of Hu sheep.These results provide a basis for further study on the function of SREBP1 gene in Hu sheep.

关 键 词：SREBP1 克隆 序列特征 湖羊 

分 类 号：S858.26[农业科学—临床兽医学]