Analysis of vaccinia virus-host protein-protein interactions: validations of yeast two-hybrid screenings.
Analysis of vaccinia virus-host protein-protein interactions: validations of yeast two-hybrid screenings.
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DOI：
10.1021/pr900491n
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发表时间：
2009-09
影响因子：
4.4
通讯作者：
McFadden, Grant
中科院分区：
文献类型：
--
作者：
Zhang, Leiliang;Villa, Nancy Y.;Rahman, Masmudur M.;Smallwood, Sherin;Shattuck, Donna;Neff, Chris;Dufford, Max;Lanchbury, Jerry S.;LaBaer, Joshua;McFadden, Grant
牛痘病毒是一种大的双链DNA病毒，是正痘病毒属的原型，正痘病毒属包括几种人类致病性痘病毒，如猴痘病毒和天花病毒。在这里，我们报告了一个全面的酵母双杂交（Y2H）筛选牛痘和人类蛋白质之间的蛋白质-蛋白质相互作用。在33种病毒蛋白中，共检测到109种新的牛痘病毒-人蛋白相互作用。为了验证这些相互作用的子集，我们使用Gateway质粒克隆系统构建了牛痘病毒株WR的ORFeome文库。通过在多种表达系统中共表达选定的牛痘和宿主蛋白质，我们发现在牛痘和人蛋白质之间鉴定的Y2H命中中的至少17个可以通过使用GST下拉测定的独立方法验证，代表了所检查的Y2H命中的63%验证率（17/27）。由于克隆的ORF可以方便地从入口载体转移到各种目的表达载体，牛痘ORFeome文库将是未来高通量功能蛋白质组学实验的有用资源。报道了一种在牛痘病毒蛋白和人蛋白之间进行的全面的酵母双杂交筛选。该筛选鉴定了涉及33种牛痘蛋白的109个命中。通过Gateway克隆系统构建了牛痘病毒ORFeome文库，并用于验证选定的相互作用。在27个检测的牛痘-人命中中，至少有17个通过GST-下拉测定被证实为阳性。一些有趣的互动进行了讨论。
Vaccinia virus, a large double-stranded DNA virus, is the prototype of the Orthopoxvirus genus, which includes several pathogenic poxviruses of humans, such as monkeypox virus and variola virus. Here, we report a comprehensive yeast two-hybrid (Y2H) screening for the protein−protein interactions between vaccinia and human proteins. A total of 109 novel vaccinia−human protein interactions were detected among 33 viral proteins. To validate subsets of those interactions, we constructed an ORFeome library of vaccinia virus strain WR using the Gateway plasmid cloning system. By co-expressing selected vaccinia and host proteins in a variety of expression systems, we found that at least 17 of the Y2H hits identified between vaccinia and human proteins can be verified by independent methods using GST pull-down assays, representing a 63% validation rate for the Y2H hits examined (17/27). Because the cloned ORFs are conveniently transferable from the entry vectors to various destination expression vectors, the vaccinia ORFeome library will be a useful resource for future high-throughput functional proteomic experiments. A comprehensive yeast two-hybrid screen between vaccinia virus proteins and human proteins is reported. This screen identified 109 hits involving 33 vaccinia proteins. A vaccinia virus ORFeome library was constructed through Gateway cloning system and used for the validation of selected interactions. At least 17 from 27 examined vaccinia−human hits were confirmed to be positive by GST-pulldown assays. Some of the interesting interactions are discussed.
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DOI：
10.1038/nrmicro1099
发表时间：
2005-03
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1083/jcb.200208112
发表时间：
2002-12-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Uchiyama K;Jokitalo E;Kano F;Murata M;Zhang X;Canas B;Newman R;Rabouille C;Pappin D;Freemont P;Kondo H
通讯作者：
Kondo H
影响因子：
3.7
作者：
Kato, Sayuri E. M.;Condit, Richard C.;Moussatche, Nissin
通讯作者：
Moussatche, Nissin
DOI：
10.1073/pnas.0900452106
发表时间：
2009-06-02
影响因子：
11.1
作者：
Mohamed, R. Mohamed;Rahman, Masmudur M.;McFadden, Grant
通讯作者：
McFadden, Grant
DOI：
10.1073/pnas.0405759101
发表时间：
2004-10-19
影响因子：
11.1
作者：
Rubins, KH;Hensley, LE;Relman, DA
通讯作者：
Relman, DA