适于双向电泳分析的酵母胞外蛋白提取方法-生物技术通报.PDFVIP

生物技术通报 ·技术与方法· BIOTECHNOLOGY BULLETIN ( ) 2015, 31 5 :73-77 适于双向电泳分析的酵母胞外蛋白提取方法 1 2 1 1 1 杜维  朴永哲  黄玮  谷月  赵长新 （1. 大连工业大学生物工程学院，大连 116034 ；2. 大连民族学院，大连 116600） 摘 要 ： 采用了无蛋白酵母培养基培养FFC2144 酵母细胞，用硫铵沉淀法、超滤法、冻干酚提等方法提取酵母胞外蛋白， 计算3 种方法的提取率并分别用双向电泳的方法对提取到的蛋白样品进行分离，同时运用质谱鉴定分离后蛋白。其中冻干- 平衡 酚法提取后得到114 个蛋白点，提取率为73.67%， 图谱识别的蛋白点最多图谱最清晰，是研究分泌类蛋白质组学理想的分离方法。 关键词 ： 酿酒酵母 ；胞外蛋白 ；双向电泳 ；提取方法 DOI ：10.13560/ki.biotech.bull.1985.2015.05.012 - Procedure to Prepare Samples for Two dimensional Electrophoresis of Secreted Proteins from Saccharomyces cerevisiae 1 2 1 1 1 Du Wei Piao Yongzhe Huang Wei Gu Yue Zhao Changxin （1. School of Biological Engineering ，Dalian Polytechnic University ，Dalian 116034 ；2. Dalian Nationalities University ，Dalian 116600 ） Abstract: Saccraomyces cerevsiae FFC2144 was cultured in nitrogen base medium without protein. The secretory proteins of yeasts were extracted by ammonium sulfate precipitation, ultrafiltration and lyophilization-phenol extraction respectively. Extraction rates by 3 methods were calculated and the proteins were separated by two-dimensional electrophoresis. The proteins isolated were confirmed by MALDI-TOF-MS. The extraction rate by lyophilization-phenol method was 73.67% and 114 protein spots were obtained with the most protein spots and clearest electrophoretogram. Lyophilization-phenol method could be an ideal separation method for studying secretory proteomics. Key words: Saccharomyces cerevisiae ；secretory protein ；two-dimensional electrophor