酿酒酵母蛋白质双向电泳条件优化及图谱建立

Construction and Optimization of Proteomic Map of Saccharomyces cerevisiae

DOI：10.3969/j.issn.1673-1689.2014.03.002

中文关键词: 酿酒酵母 全蛋白质 双向电泳

英文关键词: Saccharomyces cerevisiae total proteins two-dimensional electrophoresis

基金项目:国家自然科学基金重点项目(31130043)

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中文摘要:

利用双向电泳技术分离酿酒酵母全蛋白质,并比较了不同样品制备方法、不同凝胶参数,以及不同染色方法对电泳结果的影响。经过各个环节的条件优化,最终获得了无明显横纵条纹,背景清晰,分辨率较高,重复性较好的酿酒酵母全蛋白质双向电泳图谱。经PD-Quest软件分析,所得图谱约分离到1 000个蛋白质点,该双向电泳图谱的构建为后续氮代谢研究奠定了基础。

英文摘要:

Total proteins of Saccharomyces cerevisiae was extracted and separated by two-dimensional electrophoresis(2-DE) in this study. Different sample preparation methods,different parameters of the gel and different staining methods were compared for the optimization of 2-DE. Finally,2D-gel without obvious vertical and horizontal stripes,with low background,high resolution and good reproducibility was obtained. Analyzed by the PD-Quest software,about 1 000 protein spots were isolated from the 2-DE protein map. The construction of 2-DE protein map provide a firm basis for the follow-up nitrogen metabolism study.

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