The ORFeome Collaboration (OC) was formed, in late
2005, to meet the need of the research community for an
unrestricted source of fully sequence-validated full-ORF human cDNA
clones in a format allowing easy transfer of the ORF sequences into
virtually any type of expression vector. One of the main goal of
the project is to provide at least one fully sequenced full-ORF
clone for each of the ~18,500 currently defined human genes.
Most of the OC targets have been generated by the Dana Farber
Cancer Institute - Centre for Cancer Systems Biology (DFCI-CCSB)
during their program to develop an extensive collection of full-ORF
clones for human protein (1,2), by transferring existing MGC full
ORF human cDNA clones into Gateway™ Entry vectors. The clones were
then full-length validated at the Welcome Trust Sanger Institute
Additional human clones are being contributed by WTSI, Harvard
Institute of Proteomics (HIP), DKFZ, and Kazusa DNA Research
Institute. New full-ORF clones are converted when required at
DFCI-CCSB and then fully sequenced at WTSI.
These new clones have several advantages on cDNAs:
- High fidelity: Each CDS was amplified for only 25 cycles with
gene-specific primers and KOD HiFi Polymerase (Novagen), greatly
minimizing the risk of PCR-induced mutations.
- Time savings: ORF clones provide a shortcut to protein
expression, allowing you to skip PCR, cloning into an expression
vector, and verifying the ends of the ORF DNA sequence.
- Ease of transfer: The Gateway® (Invitrogen) entry vectors
ensure easy transfer into prokaryotic, mammalian, viral, or insect
expression systems. For maximum flexibility, the ORF stop codon has
NB: In each of these subsets all clones on one plate share the
same vector ( OCAA, OCAB and OCAC). Details of
the vectors used can be found here and more specifically
the recombination sites for each vector may be found here.
The ORFeome collaboration has made a search tool available to help you access to a
large range of information on a particular OC clone. You will have
access to the vector, insertion sites, species.
- Rual, J.F., Hirozane-Kishikawa, T., Hao, T., Bertin, N., Li,
S., Dricot, A., Li, N., Rosenberg, J., Lamesch, P., Vidalain, P.O.
et al. (2004) Human ORFeome version 1.1: a platform for reverse
proteomics. Genome Res, 14, 2128-35.
- Lamesch, P., Li, N, Milstein, S et al. (2006) hORFeome v3.1: A
Resource of Human Open Reading Frames Covering over 10,000 Human
Genes. In preparation.