Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany.
Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
Nat Methods. 2018 Aug;15(8):598-600. doi: 10.1038/s41592-018-0045-8. Epub 2018 Jul 9.
Here we describe a C-SWAT library for high-throughput tagging of Saccharomyces cerevisiae open reading frames (ORFs). In 5,661 strains, we inserted an acceptor module after each ORF that can be efficiently replaced with tags or regulatory elements. We validated the library with targeted sequencing and tagged the proteome with bright fluorescent proteins to quantify the effect of heterologous transcription terminators on protein expression and to localize previously undetected proteins.
我们在这里描述了一个用于高通量标记酿酒酵母开放阅读框（ORF）的 C-SWAT 文库。在 5661 个菌株中，我们在每个 ORF 后插入了一个接受模块，该模块可以有效地被标签或调控元件替换。我们通过靶向测序对文库进行了验证，并使用明亮的荧光蛋白标记了蛋白质组，以量化异源转录终止子对蛋白质表达的影响，并定位以前未检测到的蛋白质。
