家蚕二分浓核病毒DNA聚合酶基因启动子P97的研究  

An Investigation on DNA Polymerase-coding Gene Promoter P97 of Bombyx mori Bidensovirus

作　　者：马瑛[1] 吕鹏[1] 张苗苗[1] 胡朝阳[1] 李国辉[1] 陈克平[1] 姚勤[1] 

机构地区：[1]江苏大学生命科学研究院,江苏镇江212013

出　　处：《蚕业科学》2013年第4期716-722,共7页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31272507);国家重点基础研究发展计划"973"项目(No.2012CB114600)

摘　　要：家蚕浓核病毒2型(BmDNV-2)已被正式命名为家蚕二分浓核病毒(BmBDV),该病毒致使家蚕罹患浓核病。BmBDV是目前已知唯一能够编码DNA聚合酶的ssDNA病毒,DNA聚合酶的表达对该病毒复制至关重要。使用双荧光素酶报告系统检测BmBDV DNA聚合酶基因启动子P97的活性以及病毒潜在的调控蛋白或宿主的互作蛋白对P97的调控作用。BmBDVDNA聚合酶翻译起始位点上游286 nt序列具有启动子活性,但其活性比病毒非结构蛋白NS1基因启动子P5的活性弱。共转染瞬时表达病毒的NS1蛋白使P97启动子活性降低,而病毒的结构蛋白VP则可提高P97启动子活性;宿主家蚕的转录因子Twist蛋白的表达能够提高P97启动子的活性,但其调控作用较小。Bombyx mori densovirus 2（BmDNV-2） has been officially named Bombyx mori bidensovirus（BmBDV）.It causes densonucleosis in Bombyx mori.It is the only known ssDNA virus which encodes its own DNA polymerase so far.Expression of DNA polymerase is critical for BmBDV replication.In this study,dual-luciferase reporter assay system was used to determine the activity of DNA polymerase promoter P97 and the regulatory effect of potential viral regulatory proteins or host interacting proteins on P97.It was found that the 286 nt sequence upstream of translational initiation site of BmBDV DNA polymerase has promoter activity.Activity of P97 promoter was weaker than that of BmBDV non-structural protein NS1 gene promoter P5.After co-transfection,transiently expressed NS1 protein down-regulated the activity of P97 promoter,while structural protein VP up-regulated the activity of P97 promoter.Expression of transcriptional factor Twist protein of Bombyx mori could up-regulate the activity of P97 promoter,but the influence is quite weak.

关 键 词：家蚕二分浓核病毒 DNA聚合酶 启动子P97 活性 调控因子 

分 类 号：S884.5[农业科学—特种经济动物饲养]