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' terminus. Next, we determined the localization of 4,431 proteins, corresponding to approximately 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.
Subject(s)
Gene Expression Regulation, Fungal/drug effects , Gene Library , Gene Order/genetics , Open Reading Frames/genetics , Schizosaccharomyces pombe Proteins/genetics , Schizosaccharomyces/genetics , Antifungal Agents/pharmacology , Fatty Acids, Unsaturated/pharmacology , Genes, Fungal , Internet , Karyopherins/antagonists & inhibitors , Karyopherins/genetics , Luminescent Proteins/genetics , Proteomics/methods , Receptors, Cytoplasmic and Nuclear/antagonists & inhibitors , Receptors, Cytoplasmic and Nuclear/genetics , Saccharomyces cerevisiae Proteins/genetics , Exportin 1 ProteinABSTRACT
The ability of rice plants inoculated with Pyricularia oryzae (P. oryzae) to take up trace elements was studied by the radioactive multitracer technique. Among various elements, only Mn, Co, Zn, Se, Rb, Sr, Tc, and Re were found to be transferred to rice shoots from soil. The concentrations of essential elements, Mn and Zn, in the shoots of rice plants inoculated with P. oryzae were slightly higher than those in the control plant shoots, while Se, Rb, Tc, and Re showed almost the same concentrations for both the shoots.
Subject(s)
Ascomycota/pathogenicity , Oryza/metabolism , Oryza/microbiology , Trace Elements/pharmacokinetics , Biological Transport, Active , Oryza/growth & development , Plant Diseases/microbiology , Plant Shoots/metabolism , Plant Shoots/microbiology , Radioisotopes/pharmacokinetics , SoilABSTRACT
Heavy-ion irradiation is a new method of mutation breeding to produce new cultivars. We established the application of this method in rice plants to obtain mutants. Rice seeds were irradiated by C or Ne ions (135MeV/u) with a LET (linear energy transfer) of 22.7 or 64.2 keV/microm, respectively. Chlorophyll-deficient mutants (CDM) segregated in M2 progeny were albino, pale-green, yellow or striped-leave phenotypes. The highest rate of CDM with C-ion irradiation, 7.31%, was obtained at 40 Gy among the doses examined. Ne-ion irradiation gave the highest rate, 11.6%, at 20 Gy. We used the RLGS (Restriction Landmark Genomic Scanning) method to analyze DNA deletion in an albino mutant genome. Not I-landmark RLGS profiles detected about 2000 spots in rice. We found that one of the polymorphic spots was strongly linked to the albino phenotypic mutant derived from deleting of a DNA fragment, and demonstrated the high ability to detect of polymorphic regions by the RLGS method.
