Proteome reference maps of Medicago truncatula embryogenic cell cultures generated from single protoplasts.
Using a combination of two-dimensional gel electrophoresis (2-DE) protein mapping and mass spectrometry (MS) analysis, we have established proteome reference maps of Medicago truncatula embryogenic tissue culture cells. The cultures were generated from single protoplasts, which provided a relatively homogeneous cell population. We used these to analyze protein expression at the globular stages of somatic embryogenesis, which is the earliest morphogenetic embryonic stage. Over 3000 proteins could reproducibly be resolved over a pI range of 4-11. Three hundred and twelve protein spots were extracted from colloidal Coomassie Blue-stained 2-DE gels and analyzed by matrix-assisted laser desorption/ionization-time of flight MS analysis and tandem MS sequencing. This enabled the identification of 169 protein spots representing 128 unique gene products using a publicly available expressed sequence tag database and the MASCOT search engine. These reference maps will be valuable for the investigation of the molecular events which occur during somatic embryogenesis in M. truncatula. The proteome reference maps and supplementary materials will be available and updated for public access at http://semele.anu.edu.au/.
Imin
Nijat
N
Australian Research Council Centre of Excellence for Integrative Legume Research, Genomic Interactions Group, Research School of Biological Sciences, Australian National University, Canberra City, ACT 2601, Australia. imin@rsbs.anu.edu.au
De Jong
Femke
F
Mathesius
Ulrike
U
van Noorden
Giel
G
Saeed
Nasir A
NA
Wang
Xin-Ding
XD
Rose
Ray J
RJ
Rolfe
Barry G
BG
D000595:Amino Acid Sequence; D019992:Databases as Topic; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D020224:Expressed Sequence Tags; D016678:Genome; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted; D013058:Mass Spectrometry; D046913:Medicago truncatula / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D020543:Proteome; D011523:Protoplasts / Q000378:metabolism; D012394:Rosaniline Dyes / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D010455:Peptides; D020033:Protein Isoforms; D020543:Proteome; D012394:Rosaniline Dyes; C048139:Coomassie blue