Improved blue, green, and red fluorescent protein tagging vectors for S. cerevisiae.
Improved blue, green, and red fluorescent protein tagging vectors for S. cerevisiae.
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DOI：
10.1371/journal.pone.0067902
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Thorn KS
中科院分区：
文献类型：
--
作者：
Lee S;Lim WA;Thorn KS
荧光蛋白融合是监测蛋白质定位和运输的有力工具。这类研究特别容易在萌芽酵母酿酒酵母中进行，因为标签可以通过同源重组轻松地引入基因组。然而，现有的酵母标记质粒并没有跟上新的和改进的荧光蛋白的发展步伐。在这里，我们构建了19种不同荧光蛋白的酵母优化版本，并测试了它们在酵母中用作融合标签。这些包括两个蓝色、七个绿色和七个红色荧光蛋白，我们已经评估了它们的亮度、光稳定性和标记蛋白的扰动。我们发现EGFP仍然是表现最好的绿色荧光蛋白，TagRFP-T和mRuby2作为红色荧光蛋白表现优于mCherry，mTagBFP2可以用作蓝色荧光蛋白标签。总之，我们构建的新标签载体为酵母标签和三色成像提供了改进的蓝色和红色荧光蛋白。
Fluorescent protein fusions are a powerful tool to monitor the localization and trafficking of proteins. Such studies are particularly easy to carry out in the budding yeast Saccharomyces cerevisiae due to the ease with which tags can be introduced into the genome by homologous recombination. However, the available yeast tagging plasmids have not kept pace with the development of new and improved fluorescent proteins. Here, we have constructed yeast optimized versions of 19 different fluorescent proteins and tested them for use as fusion tags in yeast. These include two blue, seven green, and seven red fluorescent proteins, which we have assessed for brightness, photostability and perturbation of tagged proteins. We find that EGFP remains the best performing green fluorescent protein, that TagRFP-T and mRuby2 outperform mCherry as red fluorescent proteins, and that mTagBFP2 can be used as a blue fluorescent protein tag. Together, the new tagging vectors we have constructed provide improved blue and red fluorescent proteins for yeast tagging and three color imaging.
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DOI：
10.1126/science.1168683
发表时间：
2009-05-08
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Shu X;Royant A;Lin MZ;Aguilera TA;Lev-Ram V;Steinbach PA;Tsien RY
通讯作者：
Tsien RY
影响因子：
16.6
作者：
通讯作者：
--
影响因子：
3.5
作者：
Cormack, BP;Valdivia, RH;Falkow, S
通讯作者：
Falkow, S
影响因子：
64.5
作者：
Sakaue-Sawano, Asako;Kurokawa, Hiroshi;Miyawaki, Atsushi
通讯作者：
Miyawaki, Atsushi
影响因子：
3.4
作者：
Morozova, Kateryna S.;Piatkevich, Kiryl D.;Verkhusha, Vladislav V.
通讯作者：
Verkhusha, Vladislav V.