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Transcriptional Study Of PK-15 Cells In PRV Infection And Autophagy-related Analysis

Posted on:2018-08-10Degree:MasterType:Thesis
Country:ChinaCandidate:S J ZhangFull Text:PDF
GTID:2323330533457759Subject:biology
Abstract/Summary:PDF Full Text Request
PRV is a proto-type member of the genus Herpesvirus of the family Herpes virus.Pigs are infected with pseudorabies virus,with Clinical symptoms of fever,abortion and stillbirth,which caused serious economic losses to the pig industry.Transcriptome refers to the general term for all transcripts in a cell under certain growth and development periods or physiological conditions.At present,there are few studies on transcriptome of Pseudorabies virus-infected host cells.This reserach sequenced pig kidney epithelial(PK-15)cells infected with PRV for 12 hours(P-12)and for 18 hours(P-18),and analyzed the sequencing data,including sequencing data statistics and evaluation,data quality assessment,gene Expression level and difference analysis,InterProScan function annotation,GO function analysis,KEGG pathway analysis.The results showed that:(1)Compared with the control group,12 hrs after infection with PRV,we find 18546 differentially expressed transcripts,including 17427 new transcripts,and 1119 reference genes;17255 gene transcription levels were up-regulated and 1291 gene transcription levels were down-regulated in the differentially expressed genes.Eighteen hours after infection with PRV,we find 47318 differentially expressed transcripts,including 44993 new transcripts,and 2325 reference genes;44727 gene transcription levels were up-regulated and 2591 gene transcription levels were down-regulated in the differentially expressed genes.(2)IRF3?STAT6 and PKR were verified by RT-PCR after transcriptome sequencing.The results showed that IRF3 gene was up-regulated and STAT6 and PKR genes were down-regulated.This results provide a theoretical reference for further understanding the mechanism of Pseudorabies virus infection in host cells.Autophagy is a metabolic pathway that regulates the degradation of mammalian proteins by organelles.A large number of studies have shown that autophagy is extremely important for maintaining the balance of cells and organisms in the body.The research has shown that a variety of viral infections are closely related to autophagy.The relationship between PRV and autophagy has not yet been reported.This dissertation explores whether PRV can induce autophagy.PK-15 cells were chosen as experimental materials.Autophagy-related proteins LC3 and p62 were detected at different time points after PK-15 cells infected with PRV.It shows that the expression level of p62 did not change significantly.Eight hours after infection with virus,LC3 II decreased;while after 20 hours,LC3 II increased.The results indicated that autophagy was inhibited at 8 hours after virus infection and autophagy was promoted at 20 hours after virus infection.Through the exploration of Pseudorabies virus and autophagy,it provides a useful reference for the control of PRV infection.
Keywords/Search Tags:PRV, Transcriptome, gene, RT-PCR, Autophagy
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