Petrovská, B., Jeřábková, H., Chamrád, I., Vrána, J., Lenobel, R., Uřinovská, J., Sebela, M., Doležel, J.
CYTOGENETIC AND GENOME RESEARCH
143:
78-86,
2014
Keywords:
Cell cycle, Chromatin, Flow cytometry, Gel electrophoresis, Mass spectrometry, Nuclear proteome, Protein analysis
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.
Fulltext: contact IEB authors
IEB authors: Jaroslav Doležel,
Hana Jeřábková,
Beáta Petrovská,
Jan Vrána
Čeština
English