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Nat Methods. 2018 Aug;15(8):598-600. doi: 10.1038/s41592-018-0045-8. Epub 2018 Jul 9.

Genome-wide C-SWAT library for high-throughput yeast genome tagging.

Author information

1
Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany.
2
Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
3
Genome Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
4
Cell Morphogenesis and Signal Transduction, DKFZ-ZMBH Alliance and German Cancer Research Center (DKFZ), Heidelberg, Germany.
5
Department of Biophysical Chemistry, J. Heyrovsky Institute of Physical Chemistry of the CAS, Prague, Czech Republic.
6
Division of Redox Regulation, DKFZ-ZMBH Alliance and German Cancer Research Center (DKFZ), Heidelberg, Germany.
7
Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany. a.khmelinskii@imb.de.
8
Institute of Molecular Biology (IMB), Mainz, Germany. a.khmelinskii@imb.de.
9
Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel. emmanuel.levy@weizmann.ac.il.
10
Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany. m.knop@zmbh.uni-heidelberg.de.
11
Cell Morphogenesis and Signal Transduction, DKFZ-ZMBH Alliance and German Cancer Research Center (DKFZ), Heidelberg, Germany. m.knop@zmbh.uni-heidelberg.de.

Abstract

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.

PMID:
29988096
DOI:
10.1038/s41592-018-0045-8
[Indexed for MEDLINE]

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