title: Proteomic analysis of barley cell nuclei purified by flow sorting.
abstract: Many proteins are present in the nucleus; some are involved with its structural and functional organization, some with gene expression, and some with cell division. The plant nuclear proteome has not been well explored. Its characterization requires extraction methods which minimize both the artifactual alteration of the proteins and the extent of contamination with non-nuclear proteins. The conventional multi-step fractionation procedure is both laborious and prone to contamination. Here, we describe a single-step method based on flow sorting. The method allows the separation of G1, S and G2 phase nuclei and minimizes the risk of contamination by non-nuclear proteins. Preliminary results obtained using G1 phase cell nuclei from barley root tips indicate that flow sorting coupled with a protein/peptide separation and mass spectrometry will permit a comprehensive characterization of the plant nuclear proteome.
authors: Institute of Experimental Botany, Faculty of Science, Centre of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic.
authors: Beáta
authors: B
authors: Petrovská
authors: Hana
authors: H
authors: Jeřábková
authors: Ivo
authors: I
authors: Chamrád
authors: Jan
authors: J
authors: Vrána
authors: René
authors: R
authors: Lenobel
authors: Jana
authors: J
authors: Uřinovská
authors: Marek
authors: M
authors: Sebela
authors: Jaroslav
authors: J
authors: Doležel
mesh_terms: D002467:Cell Nucleus / Q000235:genetics*; D005434:Flow Cytometry / Q000379:methods; D001467:Hordeum / Q000235:genetics*; D007399:Interphase / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics; D020543:Proteome / Q000235:genetics*; D040901:Proteomics / Q000379:methods
chemical_list: D010940:Plant Proteins; D020543:Proteome