酵母Fes1蛋白与氧化应激的关系
关键词：酵母培养；氧化处理； SDS-PAGE电泳
Yeast Fes1 protein and the relationship between oxidative stress
Abstract: The experiment use Angel yeast as the material, which was incubated by yeast culture medium (YPD medium). After that, a single yeast colony was sorted out by streak cultivation. Then, a high activity yeast strains was selected under microscope to process secondary yeast propagator. On the train to log phase yeast, respectively, with heavy metal ions (Cu+ and 25μmmol/L 1mmol/L Cd+, 2 h, 3 h, respectively 4 h and 9 h, 10 h, 11 h.) And hydrogen peroxide (H2O2 solution of different concentration) stimulation treatment on yeast, the yeast of stimulus processing after centrifugal crushing, and by using polyacrylamide gel electrophoresis (SDS-PAGE electrophoresis) on the treatment of yeast protein detection.
Key words: Yeast culture; Oxidation treatment; SDS-PAGE electrophoresis
目 录
摘 要 1
引言 1
1. 材料和方法 2
1.1 实验材料 2
1.1.1 菌株 2
1.1.2培养基和缓冲液 2
1.1.4主要仪器 4
1.2 试验方法 5
1.2.1 酵母活化 5
1.2.2 酵母菌的初筛 5
1.2.3 酵母菌的复筛 5
1.2.4 酵母菌的鉴定 6
1.2.5酵母菌的扩大培养 6
1.3酵母菌的氧化刺激 6
1.3.1用不同浓度的H2O2溶液对酵母进行刺激处理 6
1.3.2分别用铜和铬两种重金属离子溶液对酵母进行刺激处理 6
1.4酵母菌的破壁 7
1.4.1破壁条件的确定 7
1.4.2酵母菌破壁 7
1.5 SDS-PAGE电泳检测蛋白 7
2. 结果与分析 8
2.1酵母菌的活化结果 8
2.2 初筛结果 8
2.3 酵母菌的鉴定 9
2.4 酵母菌氧化刺激的结果 10
2.5 酵母菌的破壁效果 11
3. 结论及展望 13
参考文献 14
致谢 15
酵母Fes1蛋白与氧化应激的关系引言
酵母Fes1蛋白的相对分子质量为32,604Da，等电点为4.76 。酵母Fes1蛋白是Hsp70(Ssa1p)核苷酸交换因素，同时是胞质Sil1p同族体，具有防止所需的释放错误折叠蛋白Hsp70 Ub-proteasome机械系统的破坏的功能, 酵母Fes1蛋白也能够增加蛋白质丰度以应对DNA复制 的压力。