Over 8000 full-length human open reading frames (ORFs) from the Mammalian Genome Collection have been cloned into the Gateway Entry vector as described by Rual et al. (2004). These clones are now available free of charge to Health Sciences researchers as part of a partnership amongst the Huntsman Cancer Institute, HSC Core Research Facilities and individual contributing investigators. To view a list of the available clones, please see the Human ORF Collection (Excel spreadsheet). The list of clones has been arranged according to the location of the clones in 96-well arrays, however this list can be easily sorted alphabetically. Note that in some cases, ORFs are represented multiple times in the collection. For more information about the collection, go to the following website: Human ORF Collection.
The clones in this collection served as the starting points for a high-throughput yeast two-hybrid analysis whereby each pairwise combination of protein products was investigated for the ability to interact (Rual et al. 2005). The Human ORF Collection list has been annotated to indicate which clones were shown to have interactors in the two-hybrid screen(a ‘Y’ in the column labeled ‘Y2H Ints’). If a clone has been shown to participate in a two-hybrid interaction, then this is a good indication that the clone encodes a functional protein. Since some ORFs are represented multiple times in the collection, it is unclear which of the clones was used in the two-hybrid screens. Therefore, the ‘Y2H Ints’ column would show a’Y’? If the ORF was involved in a yeast two-hybrid interaction but it is unknown which clone was used for the experiment. The entire list of interactions from Rual et al. 2005 can be found as a separate worksheet in this manuscript. Additional information on how the cloning was done can be found here. Additionally, many of the genes are represented on Agilent microarrays; the corresponding Agilent probes are indicated in the column labeled ‘Agilent Probe.’
A few notes of caution:
Clones have not all been sequence verified so please check to ensure that it contains the correct gene and the correct sequence. Please be sure to forward the results of your sequence confirmation to us so that we can record those clones that are verified. Note that this is annotated in the Human ORF Collection document.
When subcloning your ORF, note that the last nucleotide of the stop codon has been deleted. Please see the Human ORF request form (Acrobat pdf) diagram for graphic details.
Clones are distributed by the core as bacterial cultures on LB + 50µg/ml spectinomycin plates (for orders of 5 or more clones, please bring your own plates). To request clones or to make inquiries, please send a message to: Human.ORF.firstname.lastname@example.org. For all clone requests, please fill out the following Human ORF request form (Acrobat pdf) and send it in your email message as a PDF document.