Expression and purification of the recombinant SALT lectin from rice (Oryza sativa L.).
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.
Branco
Alan Trindade
AT
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.
Bernabé
Renato Barroso
RB
dos Santos Ferreira
Beatriz
B
de Oliveira
Marcos Vinicius Viana
MV
Garcia
Ana Beatriz
AB
de Souza Filho
Gonçalo Apolinário
GA
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
D037121:Plant Lectins; D010940:Plant Proteins; D011994:Recombinant Proteins; C076973:SALT protein, Oryza sativa