畜牧兽医学报2017，Vol.48Issue(1)：132-139,8.DOI:10.11843/j.issn.0366-6964.2017.01.016
靶向鸭肠炎病毒NP基因RNAi对DEV增殖的影响
Effection of Targeting DEV-NP Gene on DEV Proliferation by RNA Interference
摘要
Abstract
The aim of the present study was to explore the influence of nucleocapsid protein (NP)to the proliferation of duck enteritis virus (DEV).We designed and constructed pSilencer-DEV-NP according to the DEV NP gene sequence in GenBank.After these pSilencers were transfected into duck embryo fibroblast (DEF) cells in three different ways,the expression of pSilencer DEV-NP to DEV proliferation in DEF cells was analyzed by fluorescence microscope and FQ-PCR.The results showed that the green fluorescence was observed by microscope in DEF cells transfected with pSilencer-DEV-NP-1,pSilencer-DEV-NP-2,pSilencer-DEV-NP-3 and pSilencerDEV-NP-4,respectively.The silencing efficiency of pSilencer-DEV-NP-1 to 4 on DEV proliferation were 81.10％,72.69％,77.35％ and 67.69％,respectively.FQ-PCR was used to amplify and calculate.The results showed that pSilencer-DEV-NP-1 could silence the DEV proliferation in DEF cells by three transfection ways,but the silencing efficiency was different.The silencing effect of pSilencer-DEV-NP-1 was the best at 60 h after transfection and its efficiency was 69.20 ％.The second highest efficiency occurred in the way of simultaneously DEV infection and pSilencer transfection,the best situation at 48 h was 63.79％.The worst efficiency was 52.58％ at 24 h after the operation with the method of pSilencer-DEV NP-1 transfection after DEV infection.These results suggest that the nucleocapsid protein has a certain effect on DEV proliferation,which lays a theoretical foundation for clarification of the mechanism of DEV replication and proliferation.关键词
鸭肠炎病毒/核衣壳蛋白/RNA干扰/荧光定量PCRKey words
duck enteritis virus/nucleocapsid protein/RNA interference/FQ-PCR分类
农业科技引用本文复制引用
郑敏,文明,罗乐,熊朝丽,吴良涛,华敏,万润,程振涛,周碧君,杨琦..靶向鸭肠炎病毒NP基因RNAi对DEV增殖的影响[J].畜牧兽医学报,2017,48(1):132-139,8.基金项目
国家自然科学基金项目(31260607 （）
31560703) （）
贵州省优秀青年科学人才培养计划项目(黔科合人字[2013]25号) （黔科合人字[2013]25号）
贵州省百层次创新型人才项目(黔科合人才[2016]4009号) （黔科合人才[2016]4009号）
贵州省科技创新人才团队建设项目(黔科合人才团队[2015]4016号) （黔科合人才团队[2015]4016号）
贵州大学研究生创新基金项目(研农2016022) （研农2016022）
