Abstract
Medicago truncatula leaves were used as the experimental materials. Total proteins of leaves were extracted by trichloracetic acid (TCA)-acetone method and proteins had a better separation using gel strips, forming an immobilized non-linear 3 to 10 pH gradient focusing 123,000 vhr combined with 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The gels were stained with Coomassie Brilliant Blue G-250 and digitalized gels were analyzed using the PDquest 8.0.1 software. The results indicated that 931 protein dots were detected in the gel. A technology suitable for the M. truncatula leaves protein extraction by TCA/acetone and a protocol for two-dimensional electrophoresis (2-DE) was established, which provides technical support for M. truncatula leaf proteome research. Key words: Medicago truncatula, proteome, two-dimensional polyacrylamide gel electrophoresis (2-DE), isoelectrofocusing (IEF).
Highlights
Proteomics is defined as the systematic analysis of the proteome and the protein complement of genome (Pandey and Mann, 2000)
The impressive achievements in genome and expressed sequence tag (EST) sequencing have provided a wealth of information
The information is insufficient in answering questions concerning gene function, developmental/regulatory biology, and the biochemical kinetics of life
Summary
Proteomics is defined as the systematic analysis of the proteome and the protein complement of genome (Pandey and Mann, 2000). Proteome analysis has become a powerful tool in the functional characterization of plants. Due to the availability of vast nucleotide sequence information and based on the progress achieved in sensitive and rapid protein identification by mass spectrometry, proteome approaches open up new perspectives to analyze the complex functions of model plants and crop species at different levels. Complex patterns of gene expression at the level of proteins (Celis et al, 1998). The impressive achievements in genome and expressed sequence tag (EST) sequencing have provided a wealth of information. Gene expression at the transcriptional level provides important information regarding early stage transmission from genome to cellular machinery, but mRNA levels are not always consistent with the abundance of cognate proteins (Ideker et al, 2001; Gygi et al, 1999). Protein is the final executant of life functions, and the key to understanding physiological, pathological and
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