摘要
[目的]通过RNA-Seq技术筛选不同性别滩羊肌肉生长发育相关的候选基因,初步揭示滩羊背最长肌差异形成的分子机理,为其肌肉生长发育研究提供理论依据.[方法]以相同饲养条件下的8月龄滩羊为研究对象,通过转录组测序对滩羊背最长肌肉品质进行分析比较,采用Illumina HiSeqTM4000平台进行mRNA转录组测序和分析,发掘对肌肉生长发育有差异影响的相关调控基因,并对筛选的差异基因进行RT-qPCR验证.[结果]利用RNA-Seq技术测序共筛选出37个差异表达基因,其中上调基因10个,下调基因27个.对差异基因进行GO功能注释分析结果显示,其显著富集在41个GO条目中(P＜0.05);同时KEGG富集分析表明DEGs富集在28条通路中,包括FoxO、PI3K-Akt、调控干细胞多能性、嘌呤代谢、肌动蛋白细胞骨架的调节、Notch信号、蛋白质消化吸收和钙信号等通路,进一步筛选出6个基因(KSR2、ENAH、COL9A1、IFIT1、B3GAT2和TACR1).对6个差异基因进行实时荧光定量PCR验证,结果与转录组测序表达趋势一致,说明测序结果可靠.[结论]获得的KSR2、ENAH、COL9A1、IFIT1、B3GAT2和TACR1候选基因为肌肉生长发育提供更多参考资料,从而为后续宁夏优质肉羊肌肉生长发育研究奠定理论基础.
Abstract
[Objective]The present study aims to screen candidate genes related to muscle growth and development of Tan sheep with different genders by RNA-Seq technology,and initially reveal the molecular mechanism of the formation of differences in longisone dorsi muscle of Tan sheep,which can provide theoretical basis for the study of muscle growth and development.[Method]8-month-old Tan sheep under the same feeding conditions were selected as the research objects,and transcriptome sequencing was used to analyze and compare the quality of the longest back muscle of Tan sheep.mRNA transcriptome sequencing and analysis were performed using Illumina HiSeqTM4000 platform,and relevant regulatory genes with different effects on muscle growth and development were discovered.The differential genes were verified by RT-qPCR.[Result]A total of 37 differentially expressed genes were screened by RNA-Seq technology,including 10 up-regulated genes and 27 down-regulated genes.The GO function annotation analysis of differential genes showed that they were significantly enriched in 41 GO i-tems(P＜0.05).Meanwhile,KEGG enrichment analysis showed that DEGs was enriched in 28 pathways,including FoxO,PI3K-Akt,regulation of stem cell pluripotency,purine metabolism,actin cytoskeleton regulation,Notch signaling,protein digestion and absorption,calcium signaling and other pathways.Furthermore,six genes including KSR2,ENAH,COL9A1,IFIT1,B3GAT2 and TACR1 were associ-ated with muscle growth and development.Real-time fluorescence quantitative PCR was performed to verify the expression of 6 differential genes,and the results were consistent with the expression trend of transcriptome sequencing,indicating that the sequencing results were reli-able.[Conclusion]The candidate genes KSR2,ENAH,COL9A1,IFIT1,B3GAT2 and TACR1 obtained in the study can provide more refer-ence materials for muscle growth and development,thus laying a theoretical foundation for subsequent research on muscle growth and devel-opment of high-quality meat sheep in Ningxia.
基金项目
宁夏回族自治区自然科学基金(2020AAC03082)
宁夏回族自治区农业育种专项(nxnyyz20150101)
国家科技期刊平台
NSTL
万方数据