从江香猪OB基因序列及对皮下脂肪前体细胞的调控研究  

Study on OB Gene Sequence and Regulation of Subcutaneous Fat Progenitor Cells in Congjiangxiang Pig

作　　者：朱晓锋 谢玲玲 许厚强 田念念 崔小芝 吴巧群 李雄 杨云 倪萌萌 ZHU Xiaofeng;XIE Lingling;XU Houqiang;TIAN Niannian;CUI Xiaozhi;WU Qiaoqun;LI Xiong;YANG Yun;NI Mengmeng(Guizhou University College of Animal Science,Key Laboratory of Animal Genetics,Breeding and Reproduction in the Platean Mountainous Region Ministry of Education,Guizhou Guiyang 550025,China;Determination center of breeding livestock and poultry germplasm in Guizhou,Guizhou Guiyang 550018,China;Sinan county animal husbandry development center,Guizhou Tongren 565100,China)

机构地区：[1]贵州大学动物科学学院高原山地动物遗传育种与繁殖教育部重点实验室,贵州贵阳550025 [2]贵州省种畜禽种质测定中心,贵州贵阳550018 [3]思南县畜牧发展中心,贵州铜仁565100

出　　处：《中国畜牧杂志》2020年第7期96-103,共8页Chinese Journal of Animal Science

基　　金：国家科技支撑计划项目(2015BAD03B02-3);贵州省科技合作计划项目(黔科合LH字[2017]7265号);贵州省教育厅教育部重点实验室开放项目(黔教科研发[2007]222号)。

摘　　要：本文旨在探究从江香猪肥胖基因(OB sese,OB)的结构、性质以及在脂肪沉积过程中的表达调控,为进一步以从江香猪为肥胖症动物研究模型提供基础数据。研究通过提取纯种从江香猪、苏香杂交猪和大白猪心脏、肺脏、肝脏、脾脏、肾脏、大肠、小肠、脂肪、背最长肌9个组织的总RNA,逆转录后经qRT-PCR检测不同猪种不同组织中mRNA的表达水平,扩增从江香猪OB基因全长CDS序列;测序后进行生物信息学分析,构建pEGFP-C1-OB表达载体并瞬时转染至培养的从江香猪皮下脂肪前体细胞中,检测各脂质代谢相关基因的表达规律。结果表明:OB基因在从江香猪各组织器官中均有表达,其在脂肪组织中的表达水平极显著高于其他组织;与GenBank登记的野猪(Sus Scrofa)序列相比较,存在241 bp处(T→C)和462处(G→T)2个同义突变位点;氨基酸第1位至22位为蛋白信号肽区域,同时存在一个螺旋跨膜结构,概率为84.63%。表达载体转染细胞后,OB基因上调导致神经肽Y(Neuropeptide Y,NPY)、乙酰辅酶A羧化酶A(Acetyl CoA carboxylase A,ACACA)、3-磷酸甘油酰基转移酶(Glycerol-3-phosphate acyltransferase,GPAM)的表达水平极显著提高,而二酰基甘油酰基转移酶1基因、脂蛋白脂酶、过氧化物酶体增殖激活受体γ、脂肪酸合成酶的表达水平极显著降低。This paper aims to explore the structure and nature of OBsese(OB)and its expression regulation in the process of fat deposition that to provide basic data for further studies on OBsese as an animal model of OBesity.In this study,We had extracted total RNA from different tissues of purebred Congjiangxiang pig,Suxiang hybrid pig and Norther Yorkshire.After reverse transcription.The qRT-PCR had been applied to detect mRNA expression levels in different tissues of different pig species,We also had amplified the full-length CDS sequence of the OB gene of Congjiangxiang pig.The bioinformatics analysis of CDS was performed after sequencing.In order to detected the expression rules of various genes related to lipid metabolism,We had constructed pEGFP-C1-OB expression vector and transiently transfected into cultured subcutaneous fat progenitor cells of Congjiangxiang pigs,The results showed that OB gene was expressed in all tissues and organs of Congjiangxiang pig,and it was significantly higher in adipose tissue than other tissues.Compared with the sequence of Sus Scrofa in GenBank,there are two synonymous mutation sites at 241 bp(T→C)and 462(G→T)and a helical transmembrane structure,with a prOBability of 84.63%.One protein signal peptide regions from NO.1 to NO.22 of amino acids.After transfected with the expression vector,the up-regulation of OB gene resulted in significantly increased expression levels of neuropeptide Y(NPY),acetyl-coa carboxylase A(ACACA)and glycerol-3-phosphate acyltransferase(GPAM),as well as the expression levels of diacylglyceryl transferase 1 gene,lipoprotein lipase,peroxosome proliferation-activated receptor gamma and fatty acid synthase were significantly decreased.This study preliminarily investigated the up-regulation of OBesity genes in the fat precursors under the skin of Congjiangxiang pig,which led to the changes in the expression of genes related to lipid metabolism,and established the expression map of OBesity genes in various tissues of Congjiangxiang pig,providing theoretical dat

关 键 词：肥胖基因 瘦素蛋白 脂肪酸合成酶 皮下脂肪前体细胞 表达载体 原代细胞 

分 类 号：S828.2[农业科学—畜牧学]