基于RNA-Seq技术挖掘贵州白山羊公羊背最长肌生长发育下调基因  

Excavation down-regulated genes related to growth and development of longissimus dorsi tissue of male Guizhou white goat by RNA-Seq

作　　者：安清明 王星 吴震洋 孟金柱 赵园园 AN Qingming;WANG Xing;WU Zhenyang;MENG Jinzhu;ZHAO Yuanyuan(Guizhou Key Laboratory of Biodiversity Conservation and Utilization in the Fanjing Mountain Region,College of Agriculture and Forestry Engineering and Planning,Tongren University;Gansu Key Laboratory of Herbivorous Animal Biotechnology,Gansu Agricultural University)

机构地区：[1]铜仁学院农林工程与规划学院贵州省梵净山地区生物多样性保护与利用重点实验室,贵州铜仁554300 [2]甘肃农业大学草食动物生物技术重点实验室,甘肃兰州730070

出　　处：《中国兽医学报》2021年第9期1848-1856,共9页Chinese Journal of Veterinary Science

基　　金：贵州省科技计划资助项目(黔科合支撑[2020]1Y029);铜仁市科技计划资助项目([2020]75号,[2020]128号);贵州省普通高等学校科技拔尖人才支持计划资助项目(黔教合KY字[2017]089);黔科合平台人才资助项目([2020]2003号);铜仁学院创新团队资助项目(CXTD[2020-19])。

摘　　要：旨在探讨性别对贵州白山羊肌肉生长和肉品质影响的分子遗传机制,筛选出公羊背最长肌生长发育相关的下调基因。本研究采集6只同等饲养水平条件下、2周岁龄贵州白山羊背最长肌(公、母羊各3只),然后基于RNA-Seq技术分析背最长肌RNA表达情况,筛选出差异表达下调基因,并进行GO和KEGG信号通路分析,最后选取4个差异表达下调基因进行RT-PCR验证。结果显示,在所有山羊背最长肌样本中共检测到25089个有效基因(FPKM≥1,P<0.05),共筛选出514个在公羊背最长肌中显著下调的基因,其中100个为新注释到的关联基因。对获得的514个差异表达基因mRNA进行GO功能富集分析,可主要分为生物学过程和细胞组分两大类,包含21个亚类。KEGG信号通路分析发现被注释的差异基因共参与145条信号通路,其中参与核糖体信号通路的基因最为富集,共有27个基因被注释到对应的25个核糖体蛋白上。对选取的4个差异表达下调候选基因KLF11、TNS1、SIK2和IGF1R进行验证,RT-PCR结果表明4个候选基因的mRNA表达趋势与RNA-Seq检测结果一致,且差异均比较显著(P<0.05),与测序结果一致,表明RNA-Seq测序结果可靠。综上,本研究基于RNA-Seq技术共获得贵州白山羊公羊背最长肌下调基因514个,其中100个为新发掘基因,参与核糖体信号通路的基因最为富集。初步认为筛选获得的基因可作为探究贵州白山羊肌肉生长发育的候选基因,对贵州白山羊肌肉生长发育的分子改良提供了依据。This aim of this study was to investigate the molecular genetic mechanism of muscle growth and meat quality of Guizhou white goat with different genders.The longissimus dorsi tissue of six Guizhou white goat was collected,the goat were two years old and different genders with the same feeding level,then the RNA expression in longissimus dorsi tissue was analyzed based on RNA-Seq,the differentially expressed down-regulated genes were screened out,and GO and KEGG signaling pathways analysis were performed.Finally,four differentially expressed down-regulated genes(KLF11,TNS1,SIK2 and IGF1 R)were selected out,then they were verified by RT-PCR.The results showed that a total of 25089 transcripts were obtained(FPKM≥1,P<0.05),among them,514 genes were down-regulated in longissimus doris tissue of male goat,and 100 genes were new transcripts.GO functional enrichment analysis was performed on 514 differentially expressed mRNAs,which concentrated in biological process and cellular components,including 21 subclasses.KEGG signaling pathway analysis revealed that annotated differential genes participated in 145 signaling pathways,among them,the ribosomal signaling pathways were the most enriched,including27 genes and corresponding 25 ribosomal proteins.Four down-regulated candidate genes in male goat were screened out,which were KLF11,TNS1,SIK2 and IGF1 R.RT-PCR results showed that the expression trend of the four candidate genes were consistent with RNA-Seq results,and the differences of their expression levels were significant(P<0.05),indicating that the RNA-Seq results were reliable.In conclusion,514 differentially expressed down-regulated genes in longissimus doris tissue of male Guizhou white goat were screened out based on RNA-Seq technology,of which100 genes were new transcripts,the genes involved in ribosomal signaling pathway were the most enriched.The preliminary results shows that the genes obtained from the screening may be used as candidate genes to explore the muscle growth and development of Guizhou white goa

关 键 词：贵州白山羊 RNA-Seq技术 背最长肌 生长发育 候选基因 

分 类 号：S827[农业科学—畜牧学]