牛感染1型疱疹病毒(BHV-1)可导致上呼吸道疾病,结膜炎,或生殖器疾病,给全世界的养牛业造成严重的经济损失。长链非编码RNA在BHV-1感染中的作用尚不清楚。探讨lncRNA-MSCRG.16919.1在牛疱疹病毒I型(BHV-1)感染MDBK细胞中的作用,沉默lncRNA-MSTRG.16919.1基因,并通过EdgeR软件分析测序转录组和测序数据,基因本体论(GO),京都基因和基因组百科全书(KEGG),和蛋白质的相互作用网络。使用实时定量PCR(RT-qPCR)和Western印迹来验证生物信息学分析的结果。结果表明,与NC组相比,siRNA-MSCRG.16919.1组中获得了1151个差异基因。与BHV-33h相比,获得了6586个差异表达基因。从两组共筛选出498个差异表达基因。为了验证测序的准确性,随机选择6个基因进行RT-qPCR,结果表明,所选基因的表达趋势与测序结果一致。GO富集分析表明,差异基因与核苷酸结合等生物学过程有关,酶结合,细胞周期,和胶质大分子代谢.KEGG分析丰富了378和2634个信号通路,分别,与病毒感染有关,泛素介导的蛋白质水解,磷酸肌醇代谢,凋亡,和其他代谢途径。STRING蛋白质相互作用数据库用于分析由差异基因编码的蛋白质的相互作用网络,使用Cytoscape中的degree算法筛选前20种蛋白质。结果表明,SKIV2L2、JAK2、PIK3CB、MAPK8与病毒感染有关。肿瘤坏死因子的免疫印迹分析,NF-κB,MAPK8、MAPK9和MAPK10蛋白显示lncRNA-MSCRG.16919.1参与调节这些功能蛋白的表达。本研究结果为探索lncRNA-MSTRG.16919.1在生物体中的功能和调控机制提供了基础信息,也为进一步研究病毒与宿主的相互作用提供了帮助。
Infection of cattle with bovine herpesvirus type 1 (BHV-1) can lead to upper respiratory tract disease, conjunctivitis, or genital disease and cause serious economic losses to the cattle industry worldwide. The role of long noncoding RNAs in BHV-1 infection is not well understood. To explore the role of lncRNA-MSTRG.16919.1 in bovine herpes virus type I (BHV-1) infected
MDBK cells, the lncRNA-MSTRG.16919.1 gene was silenced and sequenced transcriptome and sequencing data were analyzed by Edge R software, Gene Ontology (GO), the Kyoto Encyclopedia of Genes and Genomes (KEGG), and an interaction network of proteins. Real-time quantitative PCR (RT-qPCR) and Western blotting were used to verify the results of bioinformatic analyses. The results showed that 1151 differential genes were obtained in the siRNA-MSTRG.16919.1 group compared with an NC group. Compared with BHV-33 h, 6586 differentially expressed genes were obtained. A total of 498 differentially expressed genes were screened from the two groups. To verify the accuracy of the sequencing, six genes were randomly selected for RT-qPCR, and the results showed that the expression trend of selected genes was consistent with the sequencing results. GO enrichment analysis showed that the differential genes were related to such biological processes as nucleotide binding, enzyme binding, cell cycle, and glial macromolecule metabolism. KEGG analysis enriched 378 and 2634 signaling pathways, respectively, that were associated with virus infection, ubiquitin-mediated protein hydrolysis, phosphoinositol metabolism, apoptosis, and other metabolic pathways. The STRING protein interaction database was used to analyze the interaction network of proteins encoded by differential genes, and the degree algorithm in Cytoscape was used to screen the top 20 proteins. The results showed that SKIV2L2, JAK2, PIK3CB, and MAPK8 were related to virus infection. Western blot analysis of TNF, NF-κB, MAPK8, MAPK9, and MAPK10 proteins showed that lncRNA-MSTRG.16919.1 was involved in regulating the expression of these functional proteins. The results of this study provide basic information for exploring the function and regulatory mechanism of lncRNA-MSTRG.16919.1 in organisms and help for further studying the interaction between virus and host.