title: Approaching a complete repository of sequence-verified protein-encoding clones for Saccharomyces cerevisiae.
abstract: 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 Several yeast ORF collections are available, but their use and data interpretation can be hindered by reliance on now out-of-date annotations, the inflexible presence of N- or C-terminal tags, and/or the unknown presence of mutations introduced during the cloning process.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.
authors.affiliation: Harvard Institute of Proteomics, Harvard Medical School, Cambridge, MA 02141, USA.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Hu
authors.forename: Andreas
authors.initials: A
authors.lastname: Rolfs
authors.forename: Bhupinder
authors.initials: B
authors.lastname: Bhullar
authors.forename: Tellamraju V S
authors.initials: TV
authors.lastname: Murthy
authors.forename: Cong
authors.initials: C
authors.lastname: Zhu
authors.forename: Michael F
authors.initials: MF
authors.lastname: Berger
authors.forename: Anamaria A
authors.initials: AA
authors.lastname: Camargo
authors.forename: Fontina
authors.initials: F
authors.lastname: Kelley
authors.forename: Seamus
authors.initials: S
authors.lastname: McCarron
authors.forename: Daniel
authors.initials: D
authors.lastname: Jepson
authors.forename: Aaron
authors.initials: A
authors.lastname: Richardson
authors.forename: Jacob
authors.initials: J
authors.lastname: Raphael
authors.forename: Donna
authors.initials: D
authors.lastname: Moreira
authors.forename: Elena
authors.initials: E
authors.lastname: Taycher
authors.forename: Dongmei
authors.initials: D
authors.lastname: Zuo
authors.forename: Stephanie
authors.initials: S
authors.lastname: Mohr
authors.forename: Michael F
authors.initials: MF
authors.lastname: Kane
authors.forename: Janice
authors.initials: J
authors.lastname: Williamson
authors.forename: Andrew
authors.initials: A
authors.lastname: Simpson
authors.forename: Martha L
authors.initials: ML
authors.lastname: Bulyk
authors.forename: Edward
authors.initials: E
authors.lastname: Harlow
authors.forename: Gerald
authors.initials: G
authors.lastname: Marsischky
authors.forename: Richard D
authors.initials: RD
authors.lastname: Kolodner
authors.forename: Joshua
authors.initials: J
authors.lastname: LaBaer
mesh_terms: D001482:Base Composition; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal; D023281:Genomics / Q000379:methods*; D016366:Open Reading Frames / Q000235:genetics; D016133:Polymerase Chain Reaction; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins