马立克氏病毒1.8-kb mRNA对其上游双向启动子活性的影响  

The role of 1.8-kb mRNA in regulation of Marek's disease virus bi-directional promoter between pp38 and 1.8-kb mRNA

作　　者：丁家波[1] 李延鹏[2] 杨明[3] 李中明[3] 孙爱军[3] 崔治中[3] 

机构地区：[1]中国兽医药品监察所,北京100081 [2]中国农业科学院北京畜牧兽医研究所,北京100094 [3]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医学报》2009年第11期1369-1372,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30700596)

摘　　要：构建了针对1.8-kb mRNA基因簇潜在阅读框的RNA干扰质粒pP(1.8-kb-RNAi)。将该质粒与包含其上游双向启动子的质粒pP(pp38)-CAT和pP(1.8-kb)-CAT共转染鸡胚成纤维细胞(CEF)和MDV rMd5感染的CEF(rMd5-CEF),48 h后,通过测定转染细胞裂解液中氯霉素乙酰转移酶(CAT)的活性确定1.8-kb mRNA被干扰后对双向启动子活性的影响。结果显示,利用pP(1.8-kb-RNAi)质粒干扰1.8-kb mRNA,可以使其上游双向启动子两个方向的活性均显著下降(P<0.01),其中1.8-kb mRNA方向下跌29.5%,pp38方向下跌25.0%。本研究结果证明了1.8-kb mRNA对其上游双向启动子活性有增强作用。Marek＇s disease virus （MDV） contains a bi-directional promoters located between pp38 gene and 1.8-kb mRNA in the long inverted repeat region of the viral genome. The involvement of 1.8-kb mRNA in up-regulating the activity of these promoters was analyzed by transient expression of chloramphenicol acetyltransferase （CAT） reporter gene. A plasmid pP（1.8-kb-RNAi） was constructed to interference with 1.8-kb mRNA. Two CAT reporter plasmids, pP（pp38）-CAT and pP（1.8-kb）-CAT, were transfected with pP（1.8-kb-RNAi） into chicken embryonic fibroblast （CEF） and CEF infected with rMdS,respectively. No CAT activity was detected in uninfected CEF as ex- pected. The activity of the bi-directional promoter was significantly decreased in both directions when interfered by the plasmid pP（1.8-kb-RNAi）. 29.5% and 25.0% activity were decreased in the direction of 1.8-kb mRNA family and pp38 direction, respectively. It shows that 1.8-kb mRNA plays an important role in regulating the transcriptional activity of the bi-directional promoter.

关 键 词：马立克氏病毒(MDV) 1.8-kb MRNA 双向启动子 氯霉素乙酰转移酶(CAT) RNA干扰 

分 类 号：S852.65[农业科学—基础兽医学]