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dc.contributor.authorBechtel, Stephanie
dc.contributor.authorRosenfelder, Heiko
dc.contributor.authorDuda, Anny
dc.contributor.authorPeter Schmidt, Christian
dc.contributor.authorErnst, Ute
dc.contributor.authorWellenreuther, Ruth
dc.contributor.authorMehrle, Alexander
dc.contributor.authorSchuster, Claudia
dc.contributor.authorBahr, Andre
dc.contributor.authorBlöcker, Helmut
dc.contributor.authorHeubner, Dagmar
dc.contributor.authorHoerlein, Andreas
dc.contributor.authorMichel, Guenter
dc.contributor.authorWedler, Holger
dc.contributor.authorKöhrer, Karl
dc.contributor.authorOttenwälder, Birgit
dc.contributor.authorPoustka, Annemarie
dc.contributor.authorWiemann, Stefan
dc.contributor.authorSchupp, Ingo
dc.date.accessioned2017-01-27T09:22:08Z
dc.date.available2017-01-27T09:22:08Z
dc.date.issued2007-10-31en
dc.identifier.citationBMC Genomics. 2007 Oct 31;8(1):399en
dc.identifier.urihttp://dx.doi.org/10.1186/1471-2164-8-399en
dc.identifier.urihttp://hdl.handle.net/10033/620763
dc.description.abstractAbstract Background With the completion of the human genome sequence the functional analysis and characterization of the encoded proteins has become the next urging challenge in the post-genome era. The lack of comprehensive ORFeome resources has thus far hampered systematic applications by protein gain-of-function analysis. Gene and ORF coverage with full-length ORF clones thus needs to be extended. In combination with a unique and versatile cloning system, these will provide the tools for genome-wide systematic functional analyses, to achieve a deeper insight into complex biological processes. Results Here we describe the generation of a full-ORF clone resource of human genes applying the Gateway cloning technology (Invitrogen). A pipeline for efficient cloning and sequencing was developed and a sample tracking database was implemented to streamline the clone production process targeting more than 2,200 different ORFs. In addition, a robust cloning strategy was established, permitting the simultaneous generation of two clone variants that contain a particular ORF with as well as without a stop codon by the implementation of only one additional working step into the cloning procedure. Up to 92 % of the targeted ORFs were successfully amplified by PCR and more than 93 % of the amplicons successfully cloned. Conclusion The German cDNA Consortium ORFeome resource currently consists of more than 3,800 sequence-verified entry clones representing ORFs, cloned with and without stop codon, for about 1,700 different gene loci. 177 splice variants were cloned representing 121 of these genes. The entry clones have been used to generate over 5,000 different expression constructs, providing the basis for functional profiling applications. As a member of the recently formed international ORFeome collaboration we substantially contribute to generating and providing a whole genome human ORFeome collection in a unique cloning system that is made freely available in the community.
dc.titleThe full-ORF clone resource of the German cDNA Consortiumen
dc.typeJournal Articleen
dc.language.rfc3066enen
dc.rights.holderBechtel et al.en
dc.date.updated2015-09-04T08:28:36Zen
refterms.dateFOA2018-06-13T15:31:16Z
html.description.abstractAbstract Background With the completion of the human genome sequence the functional analysis and characterization of the encoded proteins has become the next urging challenge in the post-genome era. The lack of comprehensive ORFeome resources has thus far hampered systematic applications by protein gain-of-function analysis. Gene and ORF coverage with full-length ORF clones thus needs to be extended. In combination with a unique and versatile cloning system, these will provide the tools for genome-wide systematic functional analyses, to achieve a deeper insight into complex biological processes. Results Here we describe the generation of a full-ORF clone resource of human genes applying the Gateway cloning technology (Invitrogen). A pipeline for efficient cloning and sequencing was developed and a sample tracking database was implemented to streamline the clone production process targeting more than 2,200 different ORFs. In addition, a robust cloning strategy was established, permitting the simultaneous generation of two clone variants that contain a particular ORF with as well as without a stop codon by the implementation of only one additional working step into the cloning procedure. Up to 92 % of the targeted ORFs were successfully amplified by PCR and more than 93 % of the amplicons successfully cloned. Conclusion The German cDNA Consortium ORFeome resource currently consists of more than 3,800 sequence-verified entry clones representing ORFs, cloned with and without stop codon, for about 1,700 different gene loci. 177 splice variants were cloned representing 121 of these genes. The entry clones have been used to generate over 5,000 different expression constructs, providing the basis for functional profiling applications. As a member of the recently formed international ORFeome collaboration we substantially contribute to generating and providing a whole genome human ORFeome collection in a unique cloning system that is made freely available in the community.


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