动物医学进展2016，Vol.37Issue(10)：21-25,5.
小反刍兽疫病毒原核表达体外转录系统的建立
Establishment of an in Vitro Transcription System for Peste des Petits Ruminant Virus
摘要
Abstract
In order to initiate the type of transcription pathways for PPRV,a PPRV minireplicon was con-structed and its expression in transfected cells were studied.Backbone of pGEM3Z was used to clone the coding sequences of three genes of PPRV N,P,and L and finally named pGEM3Z-N,pGEM3Z-P,pGEM3Z-L respectively.The d-ribozyme,PPRV 5′genome promoters,3′antigenome promoters,T7 terminator,chlor-amphenicol acetyltransferase sequences were synthesized by artificial way and finally named pGEM3Z-CAT.These four plasmids were transfected into the cells by lipofectamine 2000 which infected VTF7-3 with T7 RNA polymerase.The expression of the target CAT protein was analyzed by indirect immune fluo-rescence and then by Western blot.The results showed that the cotransfection displayed that specific fluo-rescence can be observed in Vero cells and the protein can be detected by Western blot.The constructed PPRV minireplicon is able to replicate and transcribe,which provides a solid foundation for further PPRV studies by reverse genetics.关键词
小反刍兽疫病毒/微型复制子/氯霉素乙酰转移酶Key words
Peste-des-petits-ruminants virus/minireplicon/chloramphenicol acetyltransferase分类
农业科技引用本文复制引用
高华峰,赵文华,高林,杨仕标..小反刍兽疫病毒原核表达体外转录系统的建立[J].动物医学进展,2016,37(10):21-25,5.基金项目
国家自然科学基金项目 （）
