一种高效的真菌总蛋白质提取方法  

An Efficient Fungal Protein Extraction Method

作　　者：胡彬彬[1] 林连兵[1] 魏云林[1] 季秀玲[1] 张琦[1] 

机构地区：[1]昆明理工大学生命科学与技术学院,昆明650500

出　　处：《中国生物工程杂志》2013年第9期53-58,共6页China Biotechnology

基　　金：国家自然科学基金(31160016);云南省应用基础研究基金(KKSA201126005);教育部回国人员科研启动基金(KKQA201226003)资助项目

摘　　要：蛋白的提取是蛋白质组学研究的首要步骤,其质量的好坏直接影响蛋白质表达分析研究的开展及其结果的准确性。为了从深黄被孢霉M6-22中提取细胞总蛋白质并进一步进行蛋白组学等一些相关研究,采用五种已报道的蛋白质提取方法分别提取深黄被孢霉M6-22的总蛋白质,以蛋白质浓度和SDS-PAGE结果分析相结合的方法比较提取方法。结果显示,其中一种方法提取的蛋白质浓度相对较高而且蛋白质条带丰度最高,进一步对该方法进行了改良,所获得蛋白质的浓度可达约2.99mg/ml。改进后的方法操作步骤简单,蛋白质条带丰度更高。采用改进后的方法提取其他五种真菌的总蛋白质,结果显示其对其他真菌也有很好的提取效果,说明可作为一种真菌总蛋白质的高效提取方法。Protein extraction is the first step of proteomics research. Its quality directly affects the protein expression analysis studies and the accuracy of the results. In order to extract total cellular protein from Mortierella isabellina strain M6-22 for further proteomics and some other related research, five protein extraction methods were tried. Protein concentration and SDS-PAGE analysis of protein abundancy were used to evaluate the effects of these methods, and one of them was selected and modified, by which, a total protein concentration of 2.99 mg/ml was obtained. The modified method had advantage of simpler procedure than the original one and could obtain higher protein abundancy. The modified method was further used to extract the total protein of five other fungi, and the results showed that similar extraction effect could be obtained, indicating that this method can be potentially used as an efficient total protein extraction method for fungi.

关 键 词：真菌总蛋白 深黄被孢霉M6-22 SDS—PAGE 

分 类 号：Q936[生物学—微生物学]