马铃薯Y病毒CP基因RNAi载体构建与干涉效果测定  

RNAi Vector Targeting and Interference Effect Determination of PVY CP Gene

作　　者：李奇科[1,2] 陶刚[2,3] 邱又彬[1,2] 邱礽[1,2] 迟胜起[4] 朱英[2,5] 刘作易[2] 

机构地区：[1]贵州大学生命科学院,贵阳550025 [2]贵州省农业生物技术重点实验室,贵阳550006 [3]贵州省植物保护研究所,贵阳550006 [4]青岛农业大学植物保护学院,青岛266109 [5]贵州省生物技术研究所,贵阳550006

出　　处：《基因组学与应用生物学》2009年第3期460-464,共5页Genomics and Applied Biology

基　　金：973计划前期研究专项(2008CB117010);贵州省科学技术重大专项(黔科合重大专项字[2008]6009号)共同资助

摘　　要：RNA沉默(RNAi)介导的植物抗病毒研究是近年来引起广泛关注的一项植物抗病毒基因工程策略。马铃薯Y病毒(PVY)在世界范围内引起马铃薯、烟草等重要经济作物的病害,并且造成严重的经济损失。本文以PVY的外壳蛋白(CP)基因为模板扩增300bp和354bp两个片段,正向和反向分别插入植物表达载体pROKⅡ的35S启动子下游,从而构建了以PVY的CP基因为靶标的RNAi植物表达载体pROKY300,转入农杆菌EHA105中,以农杆菌渗入法在本氏烟中瞬时表达与PVYCP同源的hairpin RNA。结果表明,瞬时表达的hairpinRNA有效干涉了PVY侵染。RNAi introduced gene silence recently is wildely used as an efficient method of gene engineering plant resisting virus. PVY is one of the winners causing wildly plant diseases of economic importance such as potatoes and tobaccos in the world. In this study, we amplified two segments （300 bp and 354 bp in length） of CP gene of PVY using PCR technique. By inserting those two segments sensely and antisensely respectively into the down- stream of 35S promoter of the plant expression vector-pROK Ⅱ, we constructed the RNAi expression vector which targets to CP gene. Then the vector constructed into Agrobocterium EHA105 which has been tested by detecting the transient expression of homologous CP hairpin RNA. From those results we can get the conclusion that the transient expressed hairpin RNA of CP can efficiently prevent a infection process of PVY.

关 键 词：马铃薯Y病毒 CP基因 RNAI 载体构建 农杆菌渗入法 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]