title: Expression and purification of the recombinant SALT lectin from rice (Oryza sativa L.).
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.
authors.affiliation: Laboratório de Biotecnologia, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Avenida Alberto Lamego 2000, CEP 28013-600, Parque California, Campos dos Goytacazes, RJ, Brazil.
authors.forename: Alan Trindade
authors.initials: AT
authors.lastname: Branco
authors.forename: Renato Barroso
authors.initials: RB
authors.lastname: Bernabé
authors.forename: Beatriz
authors.initials: B
authors.lastname: dos Santos Ferreira
authors.forename: Marcos Vinicius Viana
authors.initials: MV
authors.lastname: de Oliveira
authors.forename: Ana Beatriz
authors.initials: AB
authors.lastname: Garcia
authors.forename: Gonçalo Apolinário
authors.initials: GA
authors.lastname: de Souza Filho
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006386:Hemagglutination Tests / Q000379:methods; D012275:Oryza / Q000235:genetics / Q000378:metabolism*; D037121:Plant Lectins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
chemical_list: D037121:Plant Lectins; D010940:Plant Proteins; D011994:Recombinant Proteins; C076973:SALT protein, Oryza sativa