猪瘟病毒感染猪白细胞凋亡基因表达谱变化的研究  

Expression Profiles of Apoptotic Genes of Pig Peripheral Blood Leukocytes Caused by Classical Swine Fever Virus Infection

作　　者：史子学[1] 孙金福[1] 郭焕成[1] 涂长春[1] 

机构地区：[1]军事医学科学院军事兽医研究所,长春130062

出　　处：《病毒学报》2008年第6期456-463,共8页Chinese Journal of Virology

基　　金：国家基础研究重大发展计划(973)项目子课题(2005CB523200);国家科技支撑计划(2006BAD06A03)项目资助

摘　　要：为了了解CSFV感染引起猪细胞凋亡的分子机制,利用基因芯片分析了猪感染CSFV前后基因组转录水平的变化。分离攻毒前和攻毒后猪外周血白细胞(Peripheral blood leucocyte,PBL),提取总RNA,经反转录和体外转录得到cRNA,与Affymetrix猪全基因组cDNA芯片进行杂交分析,筛选差异表达的凋亡基因。结果显示,感染猪的PBLs中共筛选到24个涉及细胞凋亡有关基因发生2倍以上变化,并对差异表达的12个宿主细胞凋亡相关基因,用荧光定量RT-PCR进行了验证。功能分析显示,这些凋亡基因的表达变化与CSFV诱导宿主细胞凋亡的机制有关。本文首次用猪全基因组芯片研究了CSFV感染诱导宿主基因的表达变化,建立了CSFV感染猪细胞凋亡相关基因的表达谱,初步阐释了CSFV感染诱导宿主细胞凋亡的分子机制。Classieal swine fever （CSF） is a contagious swine disease charactered by hemorrhagic fever and leukopenia,usually leading to substantial economic losses. To obtain a insight of leucopenia caused by CSFV infection, DNA mieroarray analyses of peripheral blood leucocytes （PBL） of the infected pigs was performed. Three health pigs were inoculated with a lethal dose of CSFV Shimen strain and their PBLs were isolated when the onset of typical clinical signs and then subjected to total RNA extraction followed by microarray analysis with Affymetrix Porcine Genome Array GeneChips. The results showed that the significant differences were observed in cellular apoptotic genes expression at 7 days post-infection （p. i. ）. The changes of the genes expression were confirmed by real time RT-PCR of some selected apoptosis-related genes. This study provided a valuable information for further investigating the molecular mechanism of apoptosis caused by CSFV infection.

关 键 词：猪瘟病毒感染 基因芯片 凋亡基因表达 

分 类 号：S852.65[农业科学—基础兽医学]