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构建统一文库:通过交换标签策略简化酵母文库的创建

One library to make them all: streamlining the creation of yeast libraries via a SWAp-Tag strategy.

作者信息

Yofe Ido, Weill Uri, Meurer Matthias, Chuartzman Silvia, Zalckvar Einat, Goldman Omer, Ben-Dor Shifra, Schütze Conny, Wiedemann Nils, Knop Michael, Khmelinskii Anton, Schuldiner Maya

机构信息

Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.

Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany.

出版信息

Nat Methods. 2016 Apr;13(4):371-378. doi: 10.1038/nmeth.3795. Epub 2016 Feb 29.

Abstract

The yeast Saccharomyces cerevisiae is ideal for systematic studies relying on collections of modified strains (libraries). Despite the significance of yeast libraries and the immense variety of available tags and regulatory elements, only a few such libraries exist, as their construction is extremely expensive and laborious. To overcome these limitations, we developed a SWAp-Tag (SWAT) method that enables one parental library to be modified easily and efficiently to give rise to an endless variety of libraries of choice. To showcase the versatility of the SWAT approach, we constructed and investigated a library of ∼1,800 strains carrying SWAT-GFP modules at the amino termini of endomembrane proteins and then used it to create two new libraries (mCherry and seamless GFP). Our work demonstrates how the SWAT method allows fast and effortless creation of yeast libraries, opening the door to new ways of systematically studying cell biology.

摘要

酿酒酵母是依靠修饰菌株(文库)集合进行系统研究的理想选择。尽管酵母文库意义重大,且有各种各样可用的标签和调控元件,但此类文库数量稀少,因为其构建极其昂贵且费力。为克服这些限制,我们开发了一种交换标签(SWAp-Tag,SWAT)方法,该方法能轻松高效地对一个亲本文库进行修饰,从而产生无数种所需文库。为展示SWAT方法的多功能性,我们构建并研究了一个约1800个菌株的文库,这些菌株在内膜蛋白的氨基末端携带SWAT-GFP模块,然后用它创建了两个新文库(mCherry和无缝GFP)。我们的工作证明了SWAT方法如何能够快速、轻松地创建酵母文库,为系统研究细胞生物学开辟了新途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ce6/4869835/0eaff6f14b70/emss-67076-f001.jpg

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