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Separation and characterization of rice proteins
Author(s) -
Tsugita Akira,
Kawakami Takao,
Uchiyama Yotaro,
Kamo Masaharu,
Miyatake Norifumi,
Nozu Yuzo
Publication year - 1994
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150150198
Subject(s) - isoelectric focusing , gel electrophoresis , immobilized ph gradient , polyacrylamide gel electrophoresis , electrophoresis , isoelectric point , chromatography , two dimensional gel electrophoresis , chemistry , molecular weight size marker , biochemistry , biology , microbiology and biotechnology , gel electrophoresis of proteins , proteomics , gene , enzyme
Rice proteins from nine tissues and one organelle (leaf, chloroplast, stem, root, germ, dark germinated seedling, seed, bran, chaff and callus) were isolated and then separated by two‐dimensional gel electrophoresis (2‐DE). The protein spots were characterized according to molecular weight, isoelectric point and partial amino‐terminal sequence. Electrophoresis was carried out by isoelectric focusing (IEF), nonequilibrium pH gradient electrophoresis (NEPHGE) and immobilized pH gradient (IPG) in the first dimension, and by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) in the second dimension. With the aid of nine marker proteins, the patterns of IEF, NEPHGE and IPG 2‐DE gels were graphically combined by computer into a single synthetic image for each tissue, respectively, and these images for the nine tissues and one organelle were again combined into a single 2‐DE image for the integrated rice protein spots. The rice 2‐DE gel image resolved 4892 proteins. About 3% of the spots are characterized by amino‐terminal sequencing.