[1]楼望淮,蒋甲福,陈素梅,等.菊花B病毒外壳蛋白互作蛋白的筛选[J].南京农业大学学报,2013,36(4):43-48.[doi:10.7685/j.issn.1000-2030.2013.04.008]
 LOU Wanghuai,JIANG Jiafu,CHEN Sumei,et al.Screening of proteins interacting with the coat protein of Chrysanthemum virus B[J].Journal of Nanjing Agricultural University,2013,36(4):43-48.[doi:10.7685/j.issn.1000-2030.2013.04.008]
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菊花B病毒外壳蛋白互作蛋白的筛选()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
36卷
期数:
2013年4期
页码:
43-48
栏目:
OA栏目
出版日期:
2013-07-15

文章信息/Info

Title:
Screening of proteins interacting with the coat protein of Chrysanthemum virus B
作者:
楼望淮 蒋甲福 陈素梅 房伟民 陈发棣 管志勇 廖园
南京农业大学园艺学院, 江苏 南京 210095
Author(s):
LOU Wanghuai JIANG Jiafu CHEN Sumei FANG Weiming CHEN Fadi GUAN Zhiyong LIAO Yuan
College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
关键词:
菊花菊花B病毒外壳蛋白E3泛素连接酶酵母双杂交
Keywords:
chrysanthemumChrysanthemum virus Bcoat proteinE3 ubiquitin ligaseyeast two-hybrid system
分类号:
S682.3
DOI:
10.7685/j.issn.1000-2030.2013.04.008
摘要:
采用PCR扩增菊花B病毒(CVB)外壳蛋白基因的开放阅读框,构建酵母双杂交系统所需的诱饵表达载体pGBKT7-CVBCP,测序验证后转化Y2HGold酵母菌。将酵母菌Y187/pGADT7-cDNA与酵母菌Y2HGold/pGBKT7-CVBCP进行交配转化,在SD/-Trp/-Leu/-His/-Ade/X-α-Gal平板上筛选蓝色阳性克隆并测序pGADT7-cDNA的插入片段。结果表明:诱饵载体表达产物对酵母无毒性作用,且对报告基因无自激活作用。筛选菊花cDNA文库初步得到了与CVB外壳蛋白相互作用的E3泛素连接酶ARIADNE-like蛋白和ATP结合蛋白,为进一步研究CVB外壳蛋白与寄主的互作机制奠定了基础。
Abstract:
To find the protein in the chrysanthemum which was interacted with Chrysanthemum virus B coat protein,this experiment used PCR to get the ORF of the Chrysanthemum virus B coat protein gene constructed vector pGBKT7-CVBCP as the bait in the yeast two-hybrid system,verified by sequencing and transformed into yeast strain Y2HGold.The yeast Y187/pGADT7-cDNA was mated with yeast Y2HGold/pGBKT7-CVBCP,then SD/-Trp/-Leu/-His/-Ade/X-α-Gal plates were used for screening of blue-positive clones whose fragments inserted in the pGADT7-cDNA were sequenced.The experiment showed that the bait protein was non-toxic to the yeast and had no self-activation function.One E3 ubiquitin ligase ARIADNE-like protein and the ATP-binding protein were interacted with the Chrysanthemum virus B coat protein and that laid the foundation of the mutual interaction mechanism between the CVB coat protein and host proteins.

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备注/Memo

备注/Memo:
收稿日期:2012-6-4;改回日期:。
基金项目:江苏省科技支撑计划项目(BE2011325);教育部新世纪优秀人才支持计划项目(NCET-10-0492);国家农业科技成果转化项目(2010GB2360063)
作者简介:楼望淮,硕士研究生。
通讯作者:陈发棣,教授,博导,主要从事菊花遗传育种与分子生物学研究,E-mail:chenfd@njau.edu.cn。
更新日期/Last Update: 1900-01-01