About the ORFeome Collaboration Resource
All OC ORF clones have been cloned to specifically contain the protein-coding part of the respective genes, intentionally omitting the 5’ and 3’ UTR sequences present in mRNAs. This allows for the expression of native or of fusion proteins, depending on the selected conditions. Suitable vector systems are widely available allowing recombinant expression of proteins in almost any biological system of choice.
Most OC ORF clones are configured without stop codons at the end of the ORF, permitting the synthesis of C-terminal fusion proteins, as well as N-terminal fusion proteins. In contrast, ORFs with termination codons permit the synthesis of native proteins, as well as N-terminal fusion proteins. The OC aims to eventually provide all its ORF clones in both formats, with and without stop codons. Information on the stop codon status is provided in the clone annotation for each respective OC ORF clone.
The majority of OC ORF clones have been generated by PCR amplification of open reading frames (ORF) from full-length cDNAs representing the respective genes. However, a number of OC clones were generated either by reverse transcriptase-PCR from primary RNA or by direct DNA synthesis and subsequent cloning of the ORF sequences. The OC clone annotation provides information on the source of individual clones, like parent cDNA clones (GenBank accession numbers and clone IDs).
Depending on the respective contributing partners, different vectors and libraries were generated now comprising the OC collection. Find information on libraries, vectors, antibiotic resistance and cloning sequences (i.e., att sites upstream and downstream of ORFs) in the individual clone annotations as well as in the global description of libraries and vectors .
The following OC participants contributed to making the OC clone collection: the Mammalian Gene Collection (MGC), Dana Farber Cancer Institute-Center for Cancer Systems Biology (DFCI-CCSB), Wellcome Trust Sanger Institute (WTSI), Source BioScience™, Virginia G. Piper Center for Personalized Diagnostics at Biodesign Institute of Arizona State University (CPD), DNA Resource Core at Harvard Medical School (DF/HCC ), German Cancer Research Center (DKFZ), GeneCopoeia, Inc., RIKEN Yokohama Institute, Kazusa DNA Research Institute (KDRI), and the I.M.A.G.E. Consortium. Additional participants are possible in the future.
The bulk of the OC targets have been generated by the DFCI-CCSB, from their program to develop an extensive collection of full-ORF clones for human proteins. DFCI-CCSB has transferred into Gateway™ Entry vectors the ORF sequences from a majority of the existing MGC full-ORF human cDNA clones. Full-length sequence validation of these clones is then conducted at WTSI or BROAD.
The bioinformatics analysis of gene targets and isolated cDNA clones as well as OC clone annotation is conducted jointly between the NCBI, Harvard Medical School and the DKFZ.
The OC has focused primarily, to date, on providing Gateway full-ORF clones for human genes. Aproximately 2000 Gateway full-ORF mouse clones, however, shall eventually also be listed in the OC collection. These mouse clones were contributed by the MGC, as a result of its effort to synthesize full-ORF Gateway clones for additional human and mouse genes.
For further information on the Gateway system and the available cloning vectors, refer to Life Technologies.