诱导提取的dsRNA粗提液对黄瓜绿斑驳花叶病毒的抑制作用  

Inhibition of crude extracts of extracted dsRNA by induction on the cucumber green mottle mosaic virus

作　　者：郑海刚[1] 陈启建[1] 

机构地区：[1]福建农林大学植物病毒研究所,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2011年第4期346-350,共5页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金项目(30770089);福建省自然科学基金项目(2009J1069)

摘　　要：在已测序的黄瓜绿斑驳花叶病毒运动蛋白基因序列的基础上,设计特异引物,扩增全长基因,将其插入到原核表达载体L4440的2个T7启动子之间,构建能诱导形成双链RNA(dsRNA)的原核表达载体L4440-MP,并转化大肠杆菌HT115(DE3),经IPTG诱导,提取了高质量的dsRNA.将提取的dsRNA对黄瓜进行抗病性鉴定,ELISA检测结果表明,提取的dsR-NA能有效地抑制黄瓜绿斑驳花叶病毒的侵染,抗病率达到40%左右.The movement protein（MP） gene of cucumber green mottle mosaic virus was cloned using specific primers designed according to the sequenced gene.Then the PCR products were inserted into the vector L4440,which contains T7 promoter sites flanking each side of the multiple cloning site（MCS） and can thus produce dsRNAs.The resulted plasmids were transformed into the Escherichia coli HT115（DE3）.After IPTG induction,dsRNAs of high quality were obtained.The effects of the dsRNAs on the disease resistence of cucumber were then evaluated.ELISA experiments showed that about 40% of plants applied with dsRNAs were virus-free.

关 键 词：黄瓜绿斑驳花叶病毒 RNA沉默 双链RNA 

分 类 号：S432.1[农业科学—植物病理学]