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.
Rual
Jean-François
JF
Center for Cancer Systems Biology and Department of Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115, USA.
Hirozane-Kishikawa
Tomoko
T
Hao
Tong
T
Bertin
Nicolas
N
Li
Siming
S
Dricot
Amélie
A
Li
Ning
N
Rosenberg
Jennifer
J
Lamesch
Philippe
P
Vidalain
Pierre-Olivier
PO
Clingingsmith
Tracey R
TR
Hartley
James L
JL
Esposito
Dominic
D
Cheo
David
D
Moore
Troy
T
Simmons
Blake
B
Sequerra
Reynaldo
R
Bosak
Stephanie
S
Doucette-Stamm
Lynn
L
Le Peuch
Christian
C
Vandenhaute
Jean
J
Cusick
Michael E
ME
Albala
Joanna S
JS
Hill
David E
DE
Vidal
Marc
M
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