特异切割马铃薯卷叶病毒复制酶基因负链的核酶研究  

CLEAVAGE OF NEGATIVE STRAND RNA OF POTATO LEAFROLL VIRUS REPLICASE GENE IN VITRO BY RIBOZYME

作　　者：杨静华[1] 哈斯阿古拉[1] 梁成罡[1] 张鹤龄[1] 

机构地区：[1]内蒙古大学生物系

出　　处：《病毒学报》1998年第2期158-164,共7页Chinese Journal of Virology

基　　金：国家自然科学基金

摘　　要：根据锤头状核酶的作用模式，设计、合成并克隆了特异性切割马铃薯卷叶病毒中国分离株（ＰＬＲＶ－Ｃｈ）复制酶基因负链ＲＮＡ的核酶序列。以体外转录的ＰＬＲＶ－Ｃｈ复制酶基因负链ＲＮＡ作为底物，与转录的核酶ＲＮＡ共同保温，以检测核酶对底物的体外切割作用。实验结果表明。A hammerhead structure ribozyme targetting to GUC at position 3293-3295 in negative strand RNA of replicase gene of potato leafroll virus chinese isolate(PLRV-Ch)was designed,synthesized and cloned in plasmid pSPT19.The negative strand RNA fragment 280bp of replicase gene as a traget sequence was cloned in pGEM-4Z.The active ribozyme RNA and the target RNA were generated by in vitro transcription with T7 RNA polymerase.In vitro cleavage reactions showed that the target sequence of replicase gene was cleaved into two expected fragments,121nt and 159nt by designed ribozyme.

关 键 词：马铃薯卷叶病毒 复制酶基因负链 核酶 体外切割 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]