1. Expression and purification of the recombinant SALT lectin from rice (Oryza sativa L.).
- Author
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Branco AT, Bernabé RB, dos Santos Ferreira B, de Oliveira MV, Garcia AB, and de Souza Filho GA
- Subjects
- Animals, Electrophoresis, Polyacrylamide Gel methods, Erythrocytes drug effects, Escherichia coli genetics, Escherichia coli metabolism, Hemagglutination Tests methods, Oryza genetics, Plant Lectins biosynthesis, Plant Lectins genetics, Plant Lectins immunology, Plant Lectins isolation & purification, Plant Proteins genetics, Plant Proteins immunology, Rabbits, Recombinant Proteins biosynthesis, Recombinant Proteins genetics, Recombinant Proteins immunology, Recombinant Proteins isolation & purification, Oryza metabolism, Plant Proteins biosynthesis, Plant Proteins isolation & purification
- Abstract
The SALT protein is a 14.5 kDa mannose-binding lectin, originally described as preferentially expressed in rice plant roots in response to NaCl stress. Recombinant SALT lectin was produced in Escherichia coli from a cDNA clone encoding protein. After isopropyl-beta-d-thiogalactopyranoside induction, the expression level achieved was 23% of the soluble protein. The recombinant agglutinin was purified by a single-step process by dialyses against a high concentrated salt solution. After purification, hemagglutination assays of rabbit erythrocytes revealed that the recombinant SALT protein is a potent agglutinin (0.078 microg ml(-1) minimal concentration). The purified recombinant lectin was also used for comparative estimation of native protein amounts in protein extracts from rice plants by Western blot assay.
- Published
- 2004
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