摘要
目的利用高通量测序技术(Illumina平台)对感染牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)的MDBK细胞进行转录组学分析,并对筛选出的差异表达基因进行功能归类及相关分子机制研究。方法用BVDV感染MDBK细胞,分别在12、24、48和72 h收集细胞,提取总RNA进行转录组学分析。通过信号通路数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)及基因功能(Gene Ontology,GO)对差异基因进行功能的注释和富集分析。结果与未接毒的细胞相比,在0～12、13～24、25～48、49～72 h分别共有差异基因56、390、487、164个。感染病毒后12与24 h、24与48 h、48与72 h分别有9、55、270个相同差异基因;经KEGG和GO分析,在感染病毒24、48、72 h后,差异基因主要富集到P53、FoxO、NF-kB和PI3K-AKT信号通路等;筛选出感染后与免疫等相关的差异基因P21、FOXO1、GABARAPL1等,主要集中在细胞周期复制、凋亡和免疫等相关信号通路上。结论本研究通过对筛选出的差异表达基因初步归类分析,为后续BVDV感染MDBK细胞并在细胞内复制的相关分子机制研究奠定了基础。
Objective To analyze the transcriptomics of MDBK cells infected with bovine viral diarrhea virus(BVDV)by high-throughput sequencing(Illumina platform),and classify the function and investigate the relevant molecular mechanism of differentially expressed genes screened.Methods MDBK cells were infected with BVDV,of which the total RNA was extracted 12,24,48 and 72 h later respectively and analyzed for transcriptomics.The differentially expressed genes were subjected to function classification and enrichment analysis by using Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO).Results There were 56,390,487 and 164 differentially expressed genes in the MDBK cells 0~12,13~24,25~48 and 49~72 h after infection as compared with those before infection respectively.However,there were 9 the same differential genes between the cells 12 and 24 h,while 55 between those 24 and 48 h,and 270 between those 48 and 72 h,after infection.KEGG and GO analyses showed the differential genes 24,48 and 72 h after infection were mainly enriched in P53,FoxO,NF-kB and PI3 K-AKT signaling pathways.The immunity-associated differential genes screened,such as P21,FOXO1 and GABARAPL1,were mainly concentrated in the signal pathways associated with cell cycle,apoptosis and immunity.Conclusion The differentially expressed genes screened were preliminarily classified,which provided a basis for molecular study on infection and intracellular replication of BVDV in MDBK cells.
作者
马莉莉
陈月季
翟军军
倪宏波
MA Li-li;CHEN Yue-ji;ZHAI Jun-jun;NI Hong-bo(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing 163319,Heilongjiang Province,China)
出处
《中国生物制品学杂志》
CAS
CSCD
2019年第3期279-285,共7页
Chinese Journal of Biologicals
基金
畜禽循环生态养殖模式示范与推广(ZY18C07)
关键词
牛病毒性腹泻病毒
转录组学
差异表达基因
免疫通路
Bovine viral diarrhea virus(BVDV)
Transcriptome
Differentially expressed genes
Immune pathway
