非洲猪瘟病毒复制相关基因E165R启动子预测及其甲基化分析

Bioinformatics Analysis and Methylation Region Prediction of African Swine Fever Virus E165R Gene Promoter

ASF(非洲猪瘟)是由ASFV(非洲猪瘟病毒)引起猪的一种烈性、急性、出血性传染病。ASFV基因组为双股线性DNA(170~190 kb),可编码150~200种病毒,已报道ASFV的E165R基因与其复制相关,E165R编码的dUTPase蛋白结构已被解析。本文旨在利用软件和数据库对基因E165R的启动子进行生物信息学分析,进而分析其甲基化。首先,针对不同毒株的E165R启动子序列进行同源进化树分析,结果显示E165R启动子序列高度保守;然后,利用软件和数据库分析E165R启动子活性区域和转录因子,发现E165R启动子含有3个活性区域,启动子含有Sp1、c-Jun和C/EBPalp等6种转录因子,在Sp1、Oct-1和C/EBPalp结合序列上含有cg甲基化位点;最后,成功预测E165R启动子甲基化区域并设计了甲基化检测引物。本研究,首次从DNA甲基化角度出发,对ASFV的E165R基因启动子进行生物信息学分析,为下一步研究DNA甲基化影响ASFV的复制打下基础。

ASF(African Swine Fever)is a severe,a cute and hemorrhagic infectious pigs’disease caused by ASFV(African Swine Fever Virus).The ASFV genome is double-stranded linear DNA(170~190 kb),which can encode more than 150~200 viral proteins.It has been reported that the E165 R gene of ASFV was related to its replication,and the structure of the protein dUTPase encoded by E165 R.This study aimed to investigate bioinformatics of the promoter of gene E165 R,and then analyzed its promoter methylation region.The E165 R promoter sequence of different ASFV strains was highly conserved.Through the prediction of promoters,such as activity analysis software and methylation software,we observed that the E165 R promoter contains 6 transcription binding factors such as Sp1,c-Jun and C/EBPalp.Importantly,cg sites cover on binding sequences of Sp1,Oct-1 and C/EBPalp.We successfully predicted methylation region of E165 R promoter.Methylation detection primer of E165 R promoter was also designed.In this study,the ASFV E165 R gene promoter was carried out for the first time from DNA methylation.It could be founded for the next step to study the influence of DNA methylation about ASFV replication.