酵母Pim1蛋白酶对二羟酸脱水酶的降解
关键词：酵母；梯度稀释；生长曲线；层析；降解
Yeast Pim1 protease to the degradation of dibasic acid dehydration
Abstract：This experiment isolated yeast strains from Anqi wine Jiuqu. They were incubated with gradient dilution using potato medium at 30℃for 24 hours. After that, the yeast was originally screened with the medium of YPD and screened twice with spotting sample. After checked and picked under microscope, the yeast strains were transferred into YPD medium and incubated at 30℃ for 24 hours at the rotating speed of 150 r/min, and the growth curve of yeast was determined. Finally, the yeast strain organisms were centrifuged for cell crush and the extraction of Yeast Pim1 protease and dibasic acid dehydration. The Pim1 proteases were primarily purified with protein separation column chromatography, and detected by polyacryamide gel electropHoresis.To conduct another column chromatography purification yeast Pim1 protease enzyme and dibasic acid dehydration. Using polyacrylamide gel electrophoresis detection of protein degradation.
Key words: Yeast; Gradient dilution; Growth curve; Chromatography; The degration
目 录
摘 要 1
引言 2
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 5
1.2 试验方法 5
1.2.1 酵母活化 5
1.2.2 酵母菌的初筛 6
1.2.3 酵母菌的鉴定 6
1.2.4 酵母菌的扩大培养 6
1.3 酵母细胞的破碎 7
1.4 酵母细胞破壁程度检测 7
1.5 Pim1蛋白酶和二羟酸脱水酶的初步分离 8
1.6 凝胶电泳检测 9
1.7 Pim1蛋白酶对二羟酸脱水酶的降解 9
1.8凝胶电泳检测Pim1蛋白酶对二羟酸脱水酶的降解 9
2. 结果与分析 10
2.1 初筛结果 10
2.2 酵母菌的纯化结果 11
2.3 酵母菌的鉴定 11
2.4 凝胶电泳检测酵母菌破壁程度 12
2.5 Pim1蛋白酶和二羟酸脱水酶的初步分离结果 12
3. 结论 14
4. 展望 14
参考文献 14
致谢 16
酵母Pim1蛋白酶对二羟酸脱水酶的降解
引言
目前，随着现代社会的发展，人们的生活压力越来越大，由于饮食规律和上班时间的冲突，人们逐渐淡化了自己的健康问题，导致人们出现各种各样的疾病。而前列腺癌（Prostate Cancer）的发病率以跃居我国男性泌尿、生殖系统恶性肿瘤的第三位[1]。Pim1蛋白酶是一种丝氨酸/苏氨酸蛋白酶，已有研究显示Pim1蛋白酶在细胞凋亡、增值、分化和肿瘤发生过程中起重要作用，并证实其在前列腺癌中有高度表达[2]。提取Pim1蛋白并对其进行分析，对研究Pim1蛋白有很重要的积极作用，以便能更好的预防和治疗疾病。
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