文档介绍：石河子大学
硕士学位论文
黄瓜芽黄突变体的蛋白组学研究
姓名:胡丹凤
申请学位级别:硕士
专业:生物化学与分子生物学
指导教师:高剑峰;李鸿彬
20100601
黄瓜芽黄突变体的蛋白组学研究
黄瓜芽黄突变体的蛋白组学研究
姓名:胡丹凤(生物化学与分子生物学)
指导教师:高剑峰教授;李鸿彬副教授
摘要
目的:本实验以正常黄瓜和芽黄突变体黄瓜的 1-6 片真叶为研究对象,采用蛋白质双向
电泳技术,对正常黄瓜与芽黄突变体黄瓜相对应叶片组织中总蛋白进行研究,检测两者
蛋白质表达中存在的差异,然后应用 MALDI-TOF-MS 技术对筛选到的差异蛋白进行鉴
定,从中寻找可能影响黄瓜黄化现象的相关蛋白质,从而为黄化突变体分子机制研究奠
定了一定基础。
方法:实验所需的正常黄瓜与突变体体黄瓜均是在相同环境下培养,依次采集第一
至第六片真叶,液氮固样后,于-70℃冰箱保存备用。采用 TCA-丙酮法分别提取正
常黄瓜与芽黄突变体黄瓜的叶片组织的总蛋白质,准确进行定量后,进行双向凝胶
电泳分析,将得到重复性较好的凝胶进行扫描后,应用 PDQuest 软件分析,寻找到
差异表达蛋白后,对其中差异表达明显的蛋白点进行 MALDI-TOF 质谱分析。
结果与结论:本实验成功建立了黄瓜芽黄突变体的双向电泳分析体系,为日后植物蛋白
质双向电泳分析实验提供了技术平台。经软件分析,正常黄瓜叶片组织中检测到1047±
38个蛋白点,突变体中为913±27。进一步分析得到了正常黄瓜与芽黄突变体黄瓜的差
异表达蛋白63个,对其中22个蛋白点进行了MALDI-TOF质谱鉴定,其中芽黄突变体中
上调表达的蛋白质有7个,正常黄瓜组织中上调表达的有6个,无明显差异的蛋白及看家
蛋白9个。

关键词:黄瓜;芽黄突变体;双向电泳; 质谱

I
黄瓜芽黄突变体的蛋白组学研究
The proteomics research of virescent mutant
of cucumber leaf
Postgraduate:Hu dan-feng(Biochemistry and Molecular Biology)
Supervisor: jian-feng ; vice- hong-bin
Abstract
Objective: The experiment was based on both leaves tissues from the first euphylla to the sixth
in normal cucumber and virescent mutant, two-dimensional electrophoresis (2-DE) were used
to investigate the total proteins of the both materials ,in order to detect the different expressed
proteins between normal cucumber and virescent mutant ,and then identify parts of proteins
using the mass spectrometry (MS) technology , and look forward to discovering proteins that
may concern with aetiolation. Accordingly, the experiment was settled the foundation of
molecule mechanism of the virescent mutant in cucumber.
Methods: Both normal and virescent mutant cucumber plants were growed under the same
leaves from the first euphylla to the sixth were collected when they were present.
Following, both the normal and the mutant samples were quick-freezing though the liquid
nitrogen, and then preserved at -70℃freezer. The total