Branco Alan Trindade, Bernabé Renato Barroso, dos Santos Ferreira Beatriz, de Oliveira Marcos Vinicius Viana, Garcia Ana Beatriz, de Souza Filho Gonçalo Apolinário
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.
Protein Expr Purif. 2004 Jan;33(1):34-8. doi: 10.1016/j.pep.2003.08.017.
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.
SALT蛋白是一种14.5 kDa的甘露糖结合凝集素，最初被描述为在水稻植株根系中优先表达以响应NaCl胁迫。重组SALT凝集素由编码该蛋白的cDNA克隆在大肠杆菌中产生。经异丙基-β-D-硫代半乳糖苷诱导后，表达水平达到可溶性蛋白的23%。通过对高浓度盐溶液进行透析的单步方法纯化重组凝集素。纯化后，对兔红细胞进行的血凝试验表明重组SALT蛋白是一种强效凝集素（最低浓度为0.078 μg ml(-1)）。纯化的重组凝集素还用于通过蛋白质印迹分析比较估计水稻植株蛋白质提取物中天然蛋白的含量。
