Access
To read this article in full you may need to log in, make a payment or gain access through a site license (see right).
Research
Nature Biotechnology 24, 841–847 (1 July 2006) | doi:10.1038/nbt1222
ORFeome cloning and global analysis of protein localization in the fission yeast Schizosaccharomyces pombe
&
Abstract
Cloning of the entire set of an organism's protein-coding open reading frames (ORFs), or 'ORFeome', is a means of connecting the genome to downstream 'omics' applications. Here we report a proteome-scale study of the fission yeast Schizosaccharomyces pombe based on cloning of the ORFeome. Taking advantage of a recombination-based cloning system, we obtained 4,910 ORFs in a form that is readily usable in various analyses. First, we evaluated ORF prediction in the fission yeast genome project by expressing each ORF tagged at the 3|[prime]| terminus. Next, we determined the localization of 4,431 proteins, corresponding to |[sim]|90% of the fission yeast proteome, by tagging each ORF with the yellow fluorescent protein. Furthermore, using leptomycin B, an inhibitor of the nuclear export protein Crm1, we identified 285 proteins whose localization is regulated by Crm1.
To read this article in full you may need to log in, make a payment or gain access through a site license (see right).
