Inhibition of BmNPV replication in Bombyx mori cell by dsRNA triggered RNA interference

作     者：XUYing ZHUChenggang JINYongfeng ZHANGYaozhou 

作者机构：BiochemistryandMolecularBiologyInstituteofZhejiangUniversityHangzhou310029China ZhejiangUniversityofScienceHangzhou310018China 

出 版 物：《Chinese Science Bulletin》 (中国科学通报)

年 卷 期：2004年第49卷第12期

页      面：1261-1266页

核心收录：

学科分类：0906[农学-兽医学] 09[农学] 

主　　题：BmNPV复制 RNAi dsRNA 抑制 RNA干涉 蚕 转染 核型多角体病毒 

摘      要：RNA interference (RNAi) causes degradation of targeted endogenous RNA in many diverse organisms. To investigate the effect of dsRNA on silkworm cells, we trans- fected three kinds of synthetic dsRNAs of 435 bp(Ap1), 300 bp(Ap2) and 399 bp(AH) in length against the various regions of BmNPV’s DNA polymerase gene and DNA helicase gene, which are indispensable for viral replication in silkworm cells by TransMessengerTM transfection Reagent. Results indicated that in the experiment where silkworm cells were infected with wild-strain BmNPV of the three dsRNAs, Ap2 and AH can effectively suppress the replication of virus, but Ap1 had no effect on the inhibition of viral replication. Ap2 and AH can reduce the infective titer of BmNPV with a peak change of approximately 3—4 logs on day 4 post-infection. The results of reverse transcript polymerase chain reaction (RT-PCR) and DNA dot blotting also indicated that the ex- pression level of the two target genes and the quantity of viral DNA both distinctly decreased under the influence of Ap2 or AH. Furthermore, using fluorescence microscopy we analyzed the distribution patterns of dsRNA. Our studies revealed that a majority of dsRNA was localized in the nu- clear periphery discontinuously after 24 h of transfection.