Effects On The Level And Function Of P53 Binding Protein 1lncRNA-2 Derive From Different Chicken Simples Of MDV Infection

Marek's Disease(MD),a highly contagious lymphoproliferative infectious Disease of chickens,is caused by Marek's Disease Virus(MDV),and causes high morbidity and mortality of chickens.It is one of the main diseases that endanger the healthy development of chicken industry.The prevention and control of MD has been mainly focused on the immunological research on the pathogenesis of MD and the development of vaccines,while there have been few reports on the prevention and control,monitoring and detection methods of MD in the chicken population during the breeding process.The solution of this problem can provide effective means for the prevention and control of MD epidemic and outbreak.The selection of MD monitoring samples should be based on the principle that there is no influence on the production performance of chickens.Therefore,the screening of non-injurant samples MD detection markers will provide a meaningful reference for the prevention and control of MD.Long non-coding RNA(lnc RNA)is a class of endogenous RNA molecules with a length of about 200 nt that do not code.p53 is a tumor suppressor and p53 binding protein-1 is a molecule that can interact with p53.Lnc RNAs related to p53 binding protein-1 were obtained by sequencing in our research group.In this study,MDV infection of 1-day-old SPF chicks was applied to collect,detect and analyze spleen and pinna tissue samples of chickens infected with MDV at different stages.(Day Post Infection,dpi)p53 binding protein 1lnc RNA-2(TP53BP1-lnc RNA-2)expression level and function,and 42 d of MDV infection of jejunum tissues for tem observation,microstructure and TP53BP1-lnc RNA-2 and p53 expression level analysis,discuss the influences of MDV infection to chicken intestinal injury,screening of MDV infection chicken candidate markers of clinical and non damaging samples,and an analysis of the function of in vitro TP53BP1-lnc RNA-2,It provides a reference for the real-time monitoring and diagnosis of MDV infection in chickens and the screening of drug targets for prevention and control.In this study,200 1-day-old SPF chicks were selected and randomly divided into the experimental group and the control group under the same environment,and were raised in isolation.In the blank control group,100 rats were given intraperitoneal injection of PBS(p H=7.4)0.5ml/m2.In the experimental group,100 chickens were intraperitoneally injected with 0.5ml of MDV BJ-1 strain(1000PFU/ strain).Spleen and pinna tissues were collected at the 1st,7th,14 th,21st,35 th and 42 nd day after infection,respectively.q RT-PCRr and Western Blot were used to detect the expression level and function of TP53BP1-lnc RNA-2 in the spleen and pinna tissues of chickens infected with MDV.42 dpi chicken jejunal tissue was collected to observe the effect of MDV infection on the microscopic structure of chicken jejunal epithelial cells,and to detect the transcriptional level of TP53BP1-lnc RNA-2 of chicken jejunal tissue.p EGFP-N1-TP53BP1-lnc RNA-2 expression vector and p Green-TP53BP1 lnc RNA-2 sh RNA knockdown vector were constructed and transfected into chicken DF-1 cell.q RT-PCR and Western Blot were used to analyze the function of chicken TP53BP1-lnc RNA-2 in vitro.After MDV infection,there was no significant change at each time point in the blank control group.With the extension of the time of MD infection,TP53BP1-lnc RNA-2 in the spleen tissues of the experimental group showed a significant increase in 1-14 dpi(p<0.05),a significant decrease in 21dpi(p<0.05),and a significant increase in p53 protein in the 21 dpi to 42 dpi(p<0.05).TP53BP1 lnc RNA-2 in the feather pulp tissues of the experimental group showed an extremely significant decrease from 14 to 42 dpi(p<0.01),and the expression of p53 protein showed an extremely significant decrease from 1 to 14dpi(p<0.01),and an extremely significant increase from 14-42 dpi(p<0.01).In the experimental group,the microvilli of jejunal epithelial cells were ruptured,nuclear chromatin was gathered,mitochondrial cristae were ruptured,and the vacuoles were formed in the mitochondria eventually.The TP53BP1-lnc RNA-2 and p53 gene transcription levels were significantly increased(p<0.01).After the successful construction of p EGFP-N1-TP53BP1-lnc RNA-2 expression vector and transfection of DF-1 cells,the TP53BP1-lnc RNA-2 transcription level was significantly increased(p<0.01),and the p53 transcription level and protein expression level were significantly increased(p<0.05).The TP53BP1-lnc RNA-2 transcription level was significantly decreased(p<0.01)after the successful construction of p Green-TP53BP1 lnc RNA-2sh RNA vector and transfection of DF-1 cells.The transcriptional level and protein expression of p53 decreased significantly(p<0.05).The results showed that the changes of TP53BP1 lnc RNA-2 transcriptional level in chickens with different dpi may be related to the pathogenesis of MD.In clinical practice,the pinna tissue can be selected as a non-invasive sample to detect the change of p53 protein to monitor the occurrence of MD in the chicken population.At 42 dpi,the transcription level of this gene in jejunum tissue was consistent with that in spleen tissue,so jejunum tissue could also be used as a candidate marker for clinical diagnosis.TP53BP1 lnc RNA-2 may affect the apoptosis,transformation and development of T lymphocytes infected with MDV by affecting the transcription,expression level and function of the tumor-related factor p53.