酵母Trr2p蛋白的提取与分离
关键词：酵母；蛋白；对数期；柱层析；SDS聚丙烯酰胺凝胶电泳法7733
Extraction and Separation of Trr2p Protein from Yeast
Abstract: This investment isolates the pure culture of yeast from Anqi yeast powder . They were activated by gradient dilution using YPD culture medium. After that, confecting YPD medium screens the yeast for the first time and adopting spread plate method to screen twice. Finally, using streak plate method to culture the yeast and then conduct microscopic examination. We will inject the yeast into YPD liquid culture medium when we can ensure the yeast was pure culture. mapped analysis and made the growth curve. Broken yeast in the log phase to extract the desired protein using ultrasound, separation the protein mixture of different molecular weight by using column chromatography, using SDS polyacrylamide gel electrophoresis to isolate identified proteins. Through the biological information of relative molecular to invest Trr2p protein, to identify the isolated protein is Trr2p protein. The results show that, at 28 ℃ after 6 -18 hours the yeast will be in logarithmic phase separation, protein has a relative molecular mass of 37KDa, consistent with the theoretical value of Trr2p.The reasuls show that: we can do primary extraction of Trr2p protein in yeast, in order to do further research.
Key words：Yeast; Protein；Log phase；Column Chromatography；SDS Polyacrylamide Gel Electrophoresis
目 录
摘 要 1
引 言 2
1.材料和方法 2
1.1 实验材料 2
1.2 培养基和缓冲液 2
1.3 试验方法 4
2.结果与结论 6
2.1 酵母菌活化与复选结果 6
2.2 酵母菌的显微镜检结果 8
2.3 酵母菌生长曲线测定结果 8
2.4 电泳检测破碎效果结果 9
2.5 SDS-PAGE结果 9
2.6 利用生物信息学得到的相关结果 10
3.结论 13
参考文献 13
致 谢 15
酵母Trr2p蛋白的提取与分离引言
Trr2p(硫氧还蛋白还原酶)广泛存在于细菌、真菌、脊椎动物以及人类等的细胞中，是NADPH依赖的含有FAD结构域的硒酶，属于吡啶核苷酸-二硫化物氧化还原酶家族成员.，通过NADPH传递电子起到还原硫氧还蛋白的功能[1]，与NADPH、硫氧还蛋白共同构成了细胞的硫氧还蛋白系统.Trr2p有许多重要的生物学功能,与人类多种疾病的发病机制有重大关联，对于Trr2p做深入研究对生物医药的发展以及延长人类寿命有重要意义[2-4]。
酵母菌是一类单细胞真菌，在自然界分布广泛，主要生长在偏酸性的潮湿的含糖环境中[5]，是人类文明史中被应用得最早的微生物之一。Trr2p是酵母菌抗氧胁迫系统的重要组成部分，是一种硒酶，Trr2p蛋白是线粒体内直接行使氧化作用的酶之一[6~10]。对于文持细胞的氧化还原状态具有重要作用，Trr2p在酵母菌中的存在有利于酵母菌的抗氧化性和对甲萘醌的耐受性[11]。
下一篇：康乃馨的切花保鲜技术研究