| 酿酒酵母FFC2146 胞内蛋白及胞外蛋白双向电泳条件优化及图谱建立 |
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| 引用本文： | 王祥余,朴永哲,翟明昌,王晓丹,程贺,赵长新.酿酒酵母FFC2146 胞内蛋白及胞外蛋白双向电泳条件优化及图谱建立[J].微生物学通报,2011,38(2):270-274. |
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| 作者姓名： | 王祥余 朴永哲 翟明昌 王晓丹 程贺 赵长新 |
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| 作者单位： | 1. 大连工业大学辽宁省发酵工程重点实验室,辽宁,大连,116034
2. 大连民族学院生命科学学院,辽宁,大连,116600 |
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| 基金项目： | 国家“十一五”科技支撑计划重点项目(No. 2007BAK36B01) |
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| 摘 要： | 通过对酿酒酵母(Saccharomyces cerevisiae)的培养基、培养条件及蛋白质提取方案的优化,建立了酿酒酵母胞外和胞内蛋白双向电泳图谱制作方法。在YNB培养基中培养20 h,经过离心取上清-超滤-冻干可得到酿酒酵母胞外蛋白质样品;用SDS缓冲液悬浮酵母细胞-煮沸-超声-增溶,得到了酿酒酵母胞内蛋白质样品。经过双向电泳分离、硝酸银染色和PDQuest图像分析可以检测到了200多种酿酒酵母胞外蛋白和500多种酿酒酵母胞内蛋白。
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| 关 键 词： | 蛋白质 双向电泳 酿酒酵母 |
| 收稿时间： | 9/2/2010 12:00:00 AM |
| 修稿时间： | 2010/11/24 0:00:00 |
Optimization and construction of the intracellular and extracellular proteomic map of FFC2146 |
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| WANG Xiang-Yu,PIAO Yong-Zhe,ZHAI Ming-Chang,WANG Xiao-Dan,CHENG He and ZHAO Chang-Xin.Optimization and construction of the intracellular and extracellular proteomic map of FFC2146[J].Microbiology,2011,38(2):270-274. |
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| Authors: | WANG Xiang-Yu PIAO Yong-Zhe ZHAI Ming-Chang WANG Xiao-Dan CHENG He and ZHAO Chang-Xin |
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| Institution: | Liaoning Key laboratory of Fermentation Technology, Dalian Polytechnic University, Dalian, Liaoning 116034, China;College of Life Science, Dalian Nationalities University, Dalian, Liaoning 116600, China;Liaoning Key laboratory of Fermentation Technology, Dalian Polytechnic University, Dalian, Liaoning 116034, China;Liaoning Key laboratory of Fermentation Technology, Dalian Polytechnic University, Dalian, Liaoning 116034, China;Liaoning Key laboratory of Fermentation Technology, Dalian Polytechnic University, Dalian, Liaoning 116034, China;Liaoning Key laboratory of Fermentation Technology, Dalian Polytechnic University, Dalian, Liaoning 116034, China |
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| Abstract: | The aim of this study was to optimize a proper method to extract intracellular and extracellular protein from Saccharomyces cerevisiae to construct the proteomic maps. Methods of protein extraction and cultivation condition were optimized. The cells were cultured in YNB medium for 20 h and cells were separated by centrifugation. The extracellular proteins in supernatants were obtained through ultra filtration-freeze drying. Cell pellets were resuspended in SDS lysis buffer. The cell suspension were boiled for 5 min, after solubilized by sonication and stored until use. The intra- or extracellular protein from S. cerevisiae were separated by 2-DE and stained with silver nitrate. The separated protein spots in gels were analyzed by PDQuest. The results showed that over 200 spots of extracellular proteins and about 500 spots of intracellular proteins had been separated by 2-DE. |
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| Keywords: | Protein Two-dimensional electrophoresis (2-DE) Saccharomyces cerevisiae |
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