Approaching a complete repository of sequence-verified protein-encoding clones for Saccharomyces cerevisiae.
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 Several yeast ORF collections are available, but their use and data interpretation can be hindered by reliance on now out-of-date annotations, the inflexible presence of N- or C-terminal tags, and/or the unknown presence of mutations introduced during the cloning process.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.
Hu
Yanhui
Y
Harvard Institute of Proteomics, Harvard Medical School, Cambridge, MA 02141, USA.
Rolfs
Andreas
A
Bhullar
Bhupinder
B
Murthy
Tellamraju V S
TV
Zhu
Cong
C
Berger
Michael F
MF
Camargo
Anamaria A
AA
Kelley
Fontina
F
McCarron
Seamus
S
Jepson
Daniel
D
Richardson
Aaron
A
Raphael
Jacob
J
Moreira
Donna
D
Taycher
Elena
E
Zuo
Dongmei
D
Mohr
Stephanie
S
Kane
Michael F
MF
Williamson
Janice
J
Simpson
Andrew
A
Bulyk
Martha L
ML
Harlow
Edward
E
Marsischky
Gerald
G
Kolodner
Richard D
RD
LaBaer
Joshua
J
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
D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins