基于表达谱芯片挖掘抗猪瘟病毒相关基因及其在不同猪瘟抗体水平下的表达差异  

Mining Anti Classic Swine Fever Virus-related Genes Based on Microarray and Analyzing Their Transcriptional Expression under Different CSF-antibody Levels

作　　者：王晓铄[1] 俞英[1] 刘会杰[1] 肖丽[1] 翟立维 敖红[2] 王楚端[1] 

机构地区：[1]中国农业大学动物科技学院,北京100193 [2]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《畜牧兽医学报》2014年第1期14-22,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家科技支撑计划(2011BAD28B01);北京市现代农业产业技术创新团队(25019126);转基因生物新品种培育重大专项(2013ZX08011-006)

摘　　要：本研究旨在筛选抗猪瘟病毒相关基因,并分析不同猪瘟抗体水平下这些相关基因在大白和长白猪中的表达差异。首先,基于表达谱芯片技术,采用Agilent猪4×44K全基因组表达谱芯片,分析4组试验材料(病毒模拟物PolyI:C转染的猪PK15细胞、甲基化酶抑制剂Aza-CdR转染的PK15细胞、PolyI:C及Aza-CdR共同转染的PK15细胞、无处理的mock细胞),获得试验组间显著性差异表达基因。进而对差异表达基因进行聚类分析和Gene Ontology(GO)分析。结果表明,存在显著差异表达的抗粘液病基因(MX1)和双链RNA依赖的蛋白激酶R基因(PKR)主要参与抗病毒相关的生物学过程。分析2个基因在不同猪瘟抗体水平下大白猪与长白猪外周血中的表达情况发现,2个基因在猪瘟高抗组中的表达量均显著高于未免疫组(P<0.05)。此外,MX1在猪瘟高抗组中的表达存在品种间差异,大白猪中的表达量极显著高于长白猪(P<0.01)。鉴于MX1和PKR基因都是干扰素通路中的重要基因,本研究结果提示,MX1和PKR基因可作为针对抗猪瘟病毒感染的重要候选基因。The present study was conducted to screen the anti-CSF virus-related genes and to analyze the genes expression under the different CSF-antibody levels in Landrace and Yorkshire.Agilent porcine 4 X 44K microarray was used to analyze and obtain the differentially expressed genes in four research groups:virus mimic PolyI:C transfected cells,DNA methytransferase inhibitor Aza-CdR transfected PK15 cells,PolyI:C and Aza-CdR transfected cells,mock cells.The differentially expressed genes were analyzed by cluster and gene ontology(GO).The results showed that significantly differentially expressed genes of myxovirus resistant(MX1) and the double-stranded-RNA-dependent protein kinase(PKR) mainly enriched in the biological process of defense response to virus.The expression levels of both genes in peripheral blood of Landrace and Yorkshire were detected under different CSF-antibody levels.The expression levels of two genes were significantly higher in both breeds with high antibody levels than that with non-immune levels(P<0.05).In addition,MX1 was highly expressed in Yorkshire peripheral blood than that in Landrace(P<0.01).Considering MX1 and PKR are important genes in interferon pathway,the results indicate that both genes can be served as potential candidate genes for defending CSFV infection.

关 键 词：猪 表达谱 差异表达基因 抗猪瘟病毒相关基因 PolyI:C Aza-CdR 

分 类 号：S858.28[农业科学—临床兽医学]