摘要
为探究肌生长抑制素(myostatin,MSTN)基因对牛肌肉发育的具体调控机制,本研究选取同一牛场健康鲁西黄牛10头,其中通过转基因技术得到的基因编辑牛MSTN-/-和同种非转基因野生型牛各5头。分别采集两组牛腿臀肌肉样品,利用IIlumina HiSeq高通量测序技术进行转录组测序分析,通过生物信息学方法比较两组样本间的差异表达基因,并进行GO和KEGG富集分析,最后利用实时荧光定量PCR验证转录组测序数据。结果显示,基因编辑型牛和野生型牛之间共检测到18071个基因。在log2|FoldChange|≥1.48条件下,筛选出406个差异表达基因,其中347个显著上调,59个显著下调。GO功能富集分析显示,MSTN基因编辑后显著影响915个功能类别(P<0.05),差异基因主要参与结合、生物系统调节、免疫系统等相关功能。KEGG通路富集分析结果共涉及211个通路,差异基因主要富集在细胞黏附分子、趋化因子信号通路、细胞因子互作等信号通路上,进一步从中筛选出可能参与细胞生长、肌肉发育的差异基因(CD14、KIT、CSF1R、FBP1、DUSP4、ULBP21、PRKCB、SPN、CHAD、SRC)。实时荧光定量PCR检测结果显示,所选差异基因表达水平与转录组表达水平一致,证明测序结果的可靠性。本研究结果表明,MSTN基因发挥作用后可以介导多个下游基因表达,从而影响相关信号通路及生物学过程;同时,所筛选出的差异表达基因可作为进一步研究骨骼肌调控机制的候选靶标。
To explore the specific regulation mechanism of myostatin(MSTN)gene on muscle development of bovine,a total of ten healthy bovine were selected from the same farm,including five MSTN-/-gene-edited bovine and five wild-type bovine,and extracted the RNA of bovine muscle tissue sample.Collecting the calf-hip muscle samples of two groups,and the transcriptome analysis were sequenced by Illumina high-throughput sequencing technology.Bioinformatics method was used to compare the differentially expressed genes between two groups,and Go and KEGG enrichment analysis were performed.Real-time quantitative PCR was used to verify the transcriptome sequencing data.A total of 18071 genes were obtained from the gene-edited bovine and wild-type bovine.Under the conditions of log2|FoldChange|≥1.48,a total of 406 significantly differentially expressed genes were screened,in which 347 genes were up-regulated and 59 genes were down-regulated.GO functional analysis results showed that total 915 functional categories were effected after MSTN gene editing(P<0.05).The differentially expressed genes were mainly involved in the binding,regulation of biological process,immune system process and other related functions.KEGG functional enrichment analysis results indicated that differential genes were involved in total of 211 pathways,mainly concentrated in cell adhesion molecule,chemokine,cytokine interaction and other signaling pathways.The differentially expressed genes might be related to cell growth and muscle development were further screened out,including CD14,KIT,CSF1R,FBP1,DUSP4,ULBP21,PRKCB,SPN,CHAD and SRC genes.Real-time quantitative PCR results showed that the expression levels of the differentially expressed genes were basically consistent with the transcriptome expression,indicating the reliability of sequencing results.The results of this study indicated that MSTN gene could mediate the expression of multiple downstream genes after exert its role,effecting the relevant signaling pathways and biological processes.The screened
作者
苗曼宁
郭益文
胡德宝
曾雨晗
李新
张林林
丁向彬
郭宏
MIAO Manning;GUO Yiwen;HU Debao;ZENG Yuhan;LI Xin;ZHANG Linlin;DING Xiangbin;GUO Hong(Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry,College of Animal Science and Veterinary Medicine,Tianjin Agricultural University,Tianjin 300384,China)
出处
《中国畜牧兽医》
CAS
北大核心
2021年第7期2271-2281,共11页
China Animal Husbandry & Veterinary Medicine
基金
高产优质转基因肉牛新品种培养(2016ZX08007-002)。
