2.50
Hdl Handle:
http://hdl.handle.net/10033/620763
Title:
The full-ORF clone resource of the German cDNA Consortium
Authors:
Bechtel, Stephanie; Rosenfelder, Heiko; Duda, Anny; Peter Schmidt, Christian; Ernst, Ute; Wellenreuther, Ruth; Mehrle, Alexander; Schuster, Claudia; Bahr, Andre; Blöcker, Helmut; Heubner, Dagmar; Hoerlein, Andreas; Michel, Guenter; Wedler, Holger; Köhrer, Karl; Ottenwälder, Birgit; Poustka, Annemarie; Wiemann, Stefan; Schupp, Ingo
Abstract:
Abstract 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.
Citation:
BMC Genomics. 2007 Oct 31;8(1):399
Issue Date:
31-Oct-2007
URI:
http://dx.doi.org/10.1186/1471-2164-8-399; http://hdl.handle.net/10033/620763
Type:
Journal Article
Appears in Collections:
Publications of the research group Chemical Biology (CBIO)

Full metadata record

DC FieldValue Language
dc.contributor.authorBechtel, Stephanieen
dc.contributor.authorRosenfelder, Heikoen
dc.contributor.authorDuda, Annyen
dc.contributor.authorPeter Schmidt, Christianen
dc.contributor.authorErnst, Uteen
dc.contributor.authorWellenreuther, Ruthen
dc.contributor.authorMehrle, Alexanderen
dc.contributor.authorSchuster, Claudiaen
dc.contributor.authorBahr, Andreen
dc.contributor.authorBlöcker, Helmuten
dc.contributor.authorHeubner, Dagmaren
dc.contributor.authorHoerlein, Andreasen
dc.contributor.authorMichel, Guenteren
dc.contributor.authorWedler, Holgeren
dc.contributor.authorKöhrer, Karlen
dc.contributor.authorOttenwälder, Birgiten
dc.contributor.authorPoustka, Annemarieen
dc.contributor.authorWiemann, Stefanen
dc.contributor.authorSchupp, Ingoen
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.en
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
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