PRV感染PK-15细胞转录组学研究及自噬相关分析

发布时间：2018-03-22 16:13

本文选题：PRV　切入点：转录组　出处：《兰州大学》2017年硕士论文　论文类型：学位论文

【摘要】：伪狂犬病毒(Pseudorabies virus,PRV)是伪狂犬病的病原体,属于疱疹病毒科,疱疹病毒属。猪感染PRV后其临床伴有发热、流产和死胎等症状,给养猪业带来严重的经济损失。转录组是指在一定的生长发育时期或生理条件下,细胞中所有转录物的总称。目前伪狂犬病毒感染宿主细胞转录组学的相关研究较少。本研究对PRV感染12h(P-12)和PRV感染18h(P-18)的猪肾上皮细胞PK-15进行转录组测序,并对测序数据进行分析,包括测序数据统计与评估、数据比对质量评估、基因表达水平及差异分析、InterProScan功能注释、GO功能分析、KEGG通路分析。研究结果表明:(1)与对照组相比,感染PRV 12h后,发现18546个显著差异表达转录本,包含17427个新转录本,注释到基因组的参考基因1119个;其中17255个基因转录水平发生上调,1291个基因转录水平发生下调。同样感染PRV18h后,发现47318个显著差异表达转录本,包含44993个新转录本,注释到基因组的参考基因2325个;其中44727个基因转录水平上调,2591个基因转录水平下调。(2)转录组测序后,对IRF3、STAT6和PKR进行RT-PCR验证,结果表明IRF3基因出现上调,STAT6,PKR基因出现下调。这些研究结果为进一步了解伪狂犬病毒感染宿主细胞的作用机制提供了理论参考。自噬是调控哺乳动物蛋白质被细胞器降解的代谢途径,大量研究表明自噬对于维持机体组织细胞和生物体内的平衡极其重要。研究表明多种病毒感染与自噬密切相关,PRV与自噬的关系目前还没有相关的报道。本研究首先探索了PRV能否诱导自噬。我们选取了猪肾上皮细胞(PK-15)作为试验材料,PK-15感染PRV后,对不同时间点的自噬相关蛋白LC3和p62进行检测,发现p62没有发生明显变化;病毒感染8h后,LC3II减少;病毒感染20h后,LC3II增加。实验结果表明,病毒感染8h时抑制自噬,病毒感染20h时促进自噬。通过对伪狂犬病毒和自噬的探究,为控制PRV感染提供了有益的参考。
[Abstract]:Pseudorabies virus (PRV) is the pathogen of pseudorabies virus (PRV), which belongs to the family Herpesviridae. After infection with PRV, pigs have clinical symptoms such as fever, abortion and stillbirth. Serious economic losses to the pig industry. Transcriptome refers to a certain period of growth or physiological conditions, At present, there are few studies on transcriptome of pseudorabies virus infected host cells. In this study, the transcriptome of porcine renal epithelial cells (PK-15) infected with PRV and PRV were sequenced and the sequencing data were analyzed. Including sequencing data statistics and evaluation, data comparison quality assessment, gene expression level and differential analysis InterProScan functional annotation go functional analysis KEGG pathway analysis. The results showed that compared with the control group, PRV infection was 12 h after infection. It was found that 18 546 differentially expressed transcripts, including 17 427 new transcripts, were annotated with 1119 reference genes in the genome, of which 17 255 genes were up-regulated and 1 221 genes were down-regulated. We found 47318 differentially expressed transcripts, including 44993 new transcripts, 2325 reference genes annotated in the genome, of which 44727 genes were up-regulated and 2591 genes were down-regulated. The transcriptome was sequenced, and IRF3mSTAT6 and PKR were verified by RT-PCR. The results showed that IRF3 gene up-regulated STAT6PKR gene down-regulation. These results provide a theoretical reference for further understanding the mechanism of pseudorabies virus infection on host cells. Autophagy is the regulation of mammalian protein stromal cells. Metabolic pathways for organ degradation, A large number of studies have shown that autophagy is extremely important in maintaining the balance between organism, tissue, cells and organisms. Studies have shown that multiple viral infections are closely related to autophagy. There is no report on the relationship between PRV and autophagy. We first explored whether PRV could induce autophagy. We selected porcine renal epithelial cells (PK-15) as experimental material to infect PRV. The detection of autophagy associated protein LC3 and p62 at different time points showed that p62 did not change significantly; the number of p62 decreased after 8h of virus infection, and the increase of LC3II increased after 20 h of virus infection. The results showed that the inhibition of autophagy was observed at 8h of viral infection. The study of pseudorabies virus and autophagy provided a useful reference for the control of PRV infection.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S852.65

【相似文献】