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Approaching a complete repository of sequence-verified protein-encoding clones for Saccharomyces cerevisiae

¹ Harvard Institute of Proteomics, Harvard Medical School, Cambridge, Massachusetts 02141, USA; ² Division of Genetics, Department of Medicine, Brigham & Women’s Hospital, Harvard Medical School, Boston, Masschusetts 02115, USA; ³ Harvard University Graduate Biophysics Program, Cambridge, Massachusetts 02138, USA; ⁴ Ludwig Institute for Cancer Research, Sao Paulo SP Brazil 01509-010; ⁵ DF/HCC DNA Resource Core, Harvard Medical School, Cambridge, Massachusetts 02141, USA; ⁶ Ludwig Institute for Cancer Research, University of California San Diego, School of Medicine, La Jolla, California 92093, USA; ⁷ Ludwig Institute for Cancer Research, New York, New York 10158, USA; ⁸ Department of Pathology, Brigham & Women’s Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA; ⁹ Harvard-MIT Division of Health Sciences & Technology (HST), Harvard Medical School, Boston, Massachusetts 02115, USA

The availability of an annotated genome sequence for the yeast Saccharomyces cerevisiae has made possible the proteome-scale study of protein function and protein–protein interactions. These studies rely on availability of cloned open reading frame (ORF) collections that can be used for cell-free or cell-based protein expression. Several yeast ORF collections are available, but their use and data interpretation can be hindered by reliance on now out-of-date annotations, the inflexible presence of N- or C-terminal tags, and/or the unknown presence of mutations introduced during the cloning process. High-throughput biochemical and genetic analyses would benefit from a "gold standard" (fully sequence-verified, high-quality) ORF collection, which allows for high confidence in and reproducibility of experimental results. Here, we describe Yeast FLEXGene, a S. cerevisiae protein-coding clone collection that covers over 5000 predicted protein-coding sequences. The clone set covers 87% of the current S. cerevisiae genome annotation and includes full sequencing of each ORF insert. Availability of this collection makes possible a wide variety of studies from purified proteins to mutation suppression analysis, which should contribute to a global understanding of yeast protein function.

E-mail Joshua_labaer{at}hms.harvard.edu; fax (617) 324-0824.

[Supplemental material is available online at www.genome.org]

Article published online before print. Article and publication date are at http://www.genome.org/cgi/doi/10.1101/gr.6037607

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