ORFeome collaboration


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 (WTSI).

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 been removed.
OCAB Human Vectors confer Spectinomycin resistance
OCAA Human Vectors confer Kanamycin resistance
OCAC Mouse Vectors confer Spectinomycin resistance

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

Search for and order clones

 

Publications

  1. 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.
  2. 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.