Approaching a complete repository of sequence-verified protein-encoding clones for Saccharomyces cerevisiae
Murthy, Tellamraju S.
Berger, Michael F.
Camargo, Anamaria A.
Kane, Michael F.
Bulyk, Martha L.
Kolodner, Richard D.
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CitationHu, Y., A. Rolfs, B. Bhullar, T. V. S. Murthy, C. Zhu, M. F. Berger, A. A. Camargo, et al. 2007. “Approaching a Complete Repository of Sequence-Verified Protein-Encoding Clones for Saccharomyces Cerevisiae.” Genome Research 17 (4): 536–43. https://doi.org/10.1101/gr.6037607.
AbstractThe 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 Several yeast ORF collections are available, but their use and data interpretation can be hindered by reliance on now out-of-date annotations, the inflexible presence of N- or C-terminal tags, and/or the unknown presence of mutations introduced during the cloning process.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.
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