CAJ | 학술논문

利用蛋白质组学研究方法对稻瘟菌的胞外蛋白进行分析,可以更好地了解该病原菌的生物学特性和致病机制.蛋白质样品制备和双向电泳,都是蛋白质组学研究的基础.采用冷冻干燥,透析结合法与三氯乙酸-脱氧胆酸钠沉淀法(TCA-DOC沉淀法)提取液体培养稻瘟菌得到的胞外蛋白,利用17 cm pH 3.0～10.0与17 cm pH 4.0～7.0两种pH梯度IPG胶条对同一样品进行第一向等电聚焦,12%SDS-PAGE凝胶进行第二向电泳,银染后用PDQuest软件(V7.4)对电泳图谱进行分析.结果表明,在上述两种提取稻瘟菌胞外蛋白的方法中,TCA-DOC沉淀法具有较好的提取效果,应用该实验方法得到的蛋白质分布在pH 4.0～7.0之间的点数达到621个,因此在双向电泳中,采用pH 4.0～7.0的胶条具有较好的分辨率.
이용단백질조학연구방법대도온균적포외단백진행분석,가이경호지료해해병원균적생물학특성화치병궤제.단백질양품제비화쌍향전영,도시단백질조학연구적기출.채용냉동간조,투석결합법여삼록을산-탈양담산납침정법(TCA-DOC침정법)제취액체배양도온균득도적포외단백,이용17 cm pH 3.0～10.0여17 cm pH 4.0～7.0량충pH제도IPG효조대동일양품진행제일향등전취초,12%SDS-PAGE응효진행제이향전영,은염후용PDQuest연건(V7.4)대전영도보진행분석.결과표명,재상술량충제취도온균포외단백적방법중,TCA-DOC침정법구유교호적제취효과,응용해실험방법득도적단백질분포재pH 4.0～7.0지간적점수체도621개,인차재쌍향전영중,채용pH 4.0～7.0적효조구유교호적분변솔.
Proteomics analysis of Magnaporthe grisea is crucial for understanding of its biological characteristics and pathogenetic mechanisms. Protein extraction and two-dimensional gel electrophoresis (2-D) are basical methods for proteomics study. In this study, extracellular proteins of rice blast fungus, M. grisea, extracted from liquid cultured infiltration were concerntrated-dialysis through freeze drying system and TCA-DOC precipitation method, respectively. The extracted proteins were separated on first dimensional gel electrophoresis using 17cm pH 3.0～10.0 and 17cm pH 4.0～7.0 IPG strips and visualized the proteins by using silver staining follow second-dimensional electrophoresis. And protein maps were analyzed by PDQuest (V7.4) software. The results showed that the electrophoresis effect of proteins prepared by TCA-DOC precipitation is better than one by freeze drying combing with dialysis;totally 621 protein spots distributed in gel of pH 4.0～7.0 were detected. And 2-D results showed that the distinguishability of protein spots on gel used pH 4.0～7.0 IPG strips showed better than one used pH 3.0～10.0 IPG strips.