靶向鸭肠炎病毒NP基因RNAi对DEV增殖的影响

doi:10.11843/j.issn.0366-6964.2017.01.016

Abstract

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-l, pSilencer-DEV-NP-2, pSilencer-DEV-NP-3 and pSilencer-DEV-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.

CLC Number:

S852.659.1

ZHENG Min, LUO Le, XIONG Chao-li, WU Liang-tao, HUA Min, WAN Run, CHENG Zhen-tao, ZHOU Bi-jun, YANG Qi, WEN Ming. Effection of Targeting DEV-NP Gene on DEV Proliferation by RNA Interference[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(1): 132-139.

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Effection of Targeting DEV-NP Gene on DEV Proliferation by RNA Interference

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