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.

Institute of Experimental Botany Faculty of Science Centre of the Region Haná for Biotechnological and Agricultural Research Olomouc Czech Republic

Citace poskytuje Crossref.org

Monoterpenoid aryl hydrocarbon receptor allosteric antagonists protect against ultraviolet skin damage in female mice

Development of modern immunization agent against bovine papillomavirus type 1 infection based on BPV1 L1 recombinant protein

Auxin Metabolite Profiling in Isolated and Intact Plant Nuclei

Molecular Characterization of Novel x-Type HMW Glutenin Subunit 1B × 6.5 in Wheat

Auxin Metabolome Profiling in the Arabidopsis Endoplasmic Reticulum Using an Optimised Organelle Isolation Protocol

Proteome Analysis of Condensed Barley Mitotic Chromosomes

Chromosome analysis and sorting

Intact cell MALDI-TOF mass spectrometric analysis of Chroococcidiopsis cyanobacteria for classification purposes and identification of possible marker proteins

Auxins and Cytokinins-The Role of Subcellular Organization on Homeostasis

UNcleProt (Universal Nuclear Protein database of barley): The first nuclear protein database that distinguishes proteins from different phases of the cell cycle

The Position of His-Tag in Recombinant OspC and Application of Various Adjuvants Affects the Intensity and Quality of Specific Antibody Response after Immunization of Experimental Mice