[1]刘随新,石 达,陈建飞,等.猪流行性腹泻病毒N蛋白核仁定位信号对宿主细胞周期的影响[J].中国预防兽医学报,2013,(03):0.
 LIU Sui-xin,SHI Da,CHEN Jian-fei,et al.Effects on host cell cycle by nucleoli localization signal peptides of porcine epidemic diarrhea virus nucleocapsid protein[J].Chinese journal of preventive veterinary medicine,2013,(03):0.
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猪流行性腹泻病毒N蛋白核仁定位信号对宿主细胞周期的影响
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2013年03期
页码:
0
栏目:
病原生物学
出版日期:
2013-03-26

文章信息/Info

Title:
Effects on host cell cycle by nucleoli localization signal peptides of porcine epidemic diarrhea virus nucleocapsid protein
作者:
刘随新1石 达1 陈建飞1 张 鑫1 时洪艳1 刘孝珍12张 莎12王 璐1 冯 力1*
(1. 中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室/猪传染病研究室,黑龙江 哈尔滨 150001; 2. 东北农业大学,黑龙江 哈尔滨 150030)
Author(s):
LIU Sui-xin1 SHI Da1 CHEN Jian-fei1 ZHANG Xin1 SHI Hong-yan1 LIU Xiao-zhen12 ZHANG Sha12 WANG Lu1 FENG Li1*
(1. Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China; 2. Northeast Agricultural University, Harbin 150030, China)
关键词:
猪流行性腹泻病毒核衣壳蛋白核仁定位信号流式细胞术宿主细胞周期
Keywords:
porcine epidemic diarrhea virus nucleocapsid protein nucleoli localization signal flow cytometry cell cycle
文献标志码:
A
摘要:
为研究猪流行性腹泻病毒(PEDV)核衣壳蛋白(N)与宿主细胞相互作用,本研究根据前期已经鉴定的PEDV N蛋白核仁定位信号序列,利用重叠延伸PCR (SOE-PCR)方法扩增出核仁定位信号序列缺失的PEDV N基因(dN),并将其与真核表达载体pAc-GFP-C1连接,构建重组质粒pAc-GFP-dN。将pAc-GFP-dN转染于Vero E6细胞中,利用激光共聚焦显微镜分析重组质粒pAc-GFP-dN表达蛋白的定位情况,并利用流式细胞仪检测转染细胞的周期变化。实验结果表明:与完整的N蛋白相比,缺失核仁定位信号的N蛋白不能定位于细胞核仁,同时丧失了使宿主细胞周期在G2/M期停滞的能力,从而揭示了N蛋白核仁定位信号对宿主细胞具有重要的影响。
Abstract:
To study the interaction between porcine epidemic diarrhea virus (PEDV) N protein and the host cell, the PEDV N gene was amplified with the deletion of nucleoli localization signal sequence by SOE-PCR and cloned into eukaryotic expression vector pAc-GFP-C1 to construct the recombinant plasmid pAc-GFP-dN. Vero E6 cells were transfected with the pAc-GFP-dN and the subcellular localization of the expressed dN protein was analyzed by confocal microscopy. The effect on cell cycle of the transfected cell was detected by flow cytometry. The results showed that the dN protein was unable to localize in the nucleus compared to the N protein as expected; However, the dN was also unable to induce the arresting on cell cycle G2/M caused by N protein. These results revealed that the nucleoli localization signal in N protein has an important function to host cell.

参考文献/References:

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更新日期/Last Update: 2013-04-10