莱芜猪肌内脂肪和皮下脂肪转录组分析  

Transcriptome Analysis of Intramuscular and Subcutaneous Fat of Laiwu Pig

作　　者：冯卉 刘天义 SALSABEEL Yousuf 苗向阳[1] FENG Hui;LIU Tianyi;SALSABEEL Yousuf;MIAO Xiangyang(Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,中国北京100193

出　　处：《生命科学研究》2023年第2期179-188,共10页Life Science Research

基　　金：转基因生物新品种培育科技重大专项(2009ZX08008-004,2008ZX08008-003);农业科技创新工程资助项目(ASTIP-IAS05);中央级公益性科研院所基本科研业务费专项资金项目(Y2016JC22,Y2018PT68)。

摘　　要：为探究莱芜猪中不同部位脂肪沉积产生差异的关键基因,以180日龄体重相近的莱芜猪为实验对象,鉴定肌内脂肪(intramuscular fat,IMF)和皮下脂肪(subcutaneous fat,SCF)的mRNA表达谱,对差异表达mRNA进行基因本体论(Gene Ontology,GO)与京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)功能富集分析,构建平均表达量前300 mRNA的蛋白质互作网络,筛选关键调控mRNA,并分析关键mRNA对应基因的序列和蛋白质结构,探究基因表达水平、结构和功能的联系,最后随机挑选6个mRNA进行实时荧光定量PCR(real-time fluorescence quantitative PCR,qRT-PCR)验证。筛选结果显示,在IMF和SCF中共鉴定出1665个差异表达的mRNA,其中888个上调,777个下调。GO结果显示,差异表达mRNA显著富集于信号通路调节的生物过程条目、生物膜组成的细胞组成条目和细胞代谢过程中酶活性调节的分子功能条目。KEGG富集结果显示,差异表达mRNA主要参与脂肪细胞生成、脂代谢、炎症反应和癌症相关的信号通路。蛋白质互作网络和基因结构特征的分析结果显示,CXCR4(C-X-C motif chemokine receptor 4)、PECAM1(platelet and endothelial cell adhesion molecule 1)、PLAU(urokinase-type plasminogen activator)、AGT(angiotensinogen)、AKT2(AKT serine/threonine kinase 2)、HSP70.2(heat shock cognate 70-kD protein,tandem duplicate 2)、LGALS3(galectin 3)、POSTN(periostin)在调控网络中处于中心位置。此外,qRT-PCR验证结果和测序结果的趋势一致,证明测序结果真实可靠。文中IMF和SCF转录组的差异分析结果提示,AGT、CXCR4、HSP70.2和PLAU基因在莱芜猪IMF沉积和脂代谢过程中发挥关键作用,可作为调控IMF的候选基因。In order to explore the hub genes for the difference of adipogenesis and lipid metabolism in different tissues of Laiwu pigs,Laiwu pigs with similar body weight at 180 days of age were used to identify the mRNA expression profiles of intramuscular fat(IMF)and subcutaneous fat(SCF).The functions of differentially expressed mRNAs were analyzed through Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).A protein-protein interaction(PPI)network of the top 300 mRNAs with a high average expression level was constructed,key regulatory mRNAs were screened,and sequence features and protein structures of the key mRNAs were analyzed.Meantime,the reliability of the sequencing data was verified by real-time fluorescence quantitative PCR(qRT-PCR)for 6 randomly selected mRNAs.A total of 1665 differentially expressed mRNAs were identified from IMF and SCF mRNA expression profiles,of which 888 were up-regulated and 777 were down-regulated.The GO results showed that the differentially expressed mRNAs were mainly involved in regulating signal pathways in biological process(BP);composing biological membranes in cellular component(CC);affecting the enzyme activity in the cell metabolism process in molecular function(MF).The KEGG enrichment results showed that the differentially expressed mRNAs were mainly involved in adipocyte production,lipid metabolism,inflammation and cancer-related signaling pathways.Through analysis of the PPI network,gene sequence and protein structure characteristics,it was found that C-X-C motif chemokine receptor 4(CXCR4),platelet and endothelial cell adhesion molecule 1(PECAM1),urokinasetype plasminogen activator(PLAU),angiotensinogen(AGT),AKT serine/threonine kinase 2(AKT2),heat shock cognate 70-kD protein,tandem duplicate 2(HSP70.2),galectin 3(LGALS3),and periostin(POSTN)were in the center of the regulatory network.The qRT-PCR verification results were consistent with the sequencing results,proving the reliability of sequencing results.By comparing the difference between IMF and SCF transc

关 键 词：脂肪代谢 肌内脂肪(IMF) 皮下脂肪(SCF) 转录组测序 PCR验证 

分 类 号：Q812[生物学—生物工程] S828.2[农业科学—畜牧学]