Human ORFeome version 1.1: a platform for reverse proteomics.
The advent of systems biology necessitates the cloning of nearly entire sets of protein-encoding open reading frames (ORFs), or ORFeomes, to allow functional studies of the corresponding proteomes. Here, we describe the generation of a first version of the human ORFeome using a newly improved Gateway recombinational cloning approach. Using the Mammalian Gene Collection (MGC) resource as a starting point, we report the successful cloning of 8076 human ORFs, representing at least 7263 human genes, as mini-pools of PCR-amplified products. These were assembled into the human ORFeome version 1.1 (hORFeome v1.1) collection. After assessing the overall quality of this version, we describe the use of hORFeome v1.1 for heterologous protein expression in two different expression systems at proteome scale. The hORFeome v1.1 represents a central resource for the cloning of large sets of human ORFs in various settings for functional proteomics of many types, and will serve as the foundation for subsequent improved versions of the human ORFeome.
Center for Cancer Systems Biology and Department of Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
Jean-François
JF
Rual
Tomoko
T
Hirozane-Kishikawa
Tong
T
Hao
Nicolas
N
Bertin
Siming
S
Li
Amélie
A
Dricot
Ning
N
Li
Jennifer
J
Rosenberg
Philippe
P
Lamesch
Pierre-Olivier
PO
Vidalain
Tracey R
TR
Clingingsmith
James L
JL
Hartley
Dominic
D
Esposito
David
D
Cheo
Troy
T
Moore
Blake
B
Simmons
Reynaldo
R
Sequerra
Stephanie
S
Bosak
Lynn
L
Doucette-Stamm
Christian
C
Le Peuch
Jean
J
Vandenhaute
Michael E
ME
Cusick
Joanna S
JS
Albala
David E
DE
Hill
Marc
M
Vidal
D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors; D023281:Genomics / Q000379:methods*; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics* / Q000502:physiology*; D040901:Proteomics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D011994:Recombinant Proteins