植物蛋白质组学华中科技大学研究生院Word文档格式.docx

1、专 业年 龄学术方向杨广笑教授生化与分子生物学49植物生化与分子生物学涂知明副教授47课程负责教师留学经历及学术专长简介：留学经历：1996.101999.03 在日本筑波大学做博士后研究1999.042005.03 在日本国立农业生物资源研究所任特别研究员学术专长：主要从事植物生物化学与分子生物学研究, 内容涉及植物功能基因组学、蛋白质组学、植物储藏蛋白结构与功能、植物激素信号传导的分子机制及其调控的基因表达、植物对逆境反应的分子机理等。课程教学目标：蛋白质组学是二十世纪九十年代中期诞生的一门新兴学科，是新世纪生命科学研究的前沿，是应用大规模蛋白质分离和识别技术研究蛋白质组及蛋白质间相互作用

2、的一门学科。本课程着重讲授植物蛋白质组学的基本原理和研究方法，同时介绍植物蛋白质组学研究领域的最新进展。通过本课程的学习，使学生了解并初步掌握植物蛋白质大规模分离、鉴定以及蛋白质间相互的研究手段和实验方法。课程大纲：（章节目录）第一章 绪论1.1 蛋白质组学概念的提出1.2 蛋白质组学与基因组学的联系与区别1.3 植物蛋白质组学的研究意义和内容1.4 植物蛋白质组学的研究现状和发展趋势第二章 蛋白质组学的研究方法2.1 概述2.2 大规模蛋白质分离技术2.3 高通量蛋白质鉴定技术第三章 植物蛋白质提取方法3.1 概述3.2 TCA-丙酮法提取植物全蛋白质技术3.3 顽拗植物蛋白质提取技术3.4

3、 木本植物蛋白质提取技术3.5 植物种子蛋白质提取技术3.6 植物细胞器蛋白质提取技术第四章 蛋白质的分离技术4.1 一维等电聚焦电泳4.2 Blue-Native凝胶技术分析植物蛋白质复合物4.3 二维SDS聚丙烯酰胺凝胶电泳4.4 胶上蛋白的检测4.5 存在问题第五章 蛋白质组研究中的图像分析与蛋白谱数据库的建立5.1 蛋白电泳图像分析系统5.2 实用软件简介5.3 二维电泳蛋白谱数据库第六章 蛋白质测序仪与蛋白质的鉴定6.1 原理6.2 蛋白质序列分析仪和质谱仪优缺点比较6.3 样品制备6.4 分析与数据检索第七章 基质辅助激光解吸电离飞行时间质谱7.1 MALDI离子源7.2 线性飞行

4、时间质量检测器7.3 反射飞行时间质量检测器7.4 样品制备方法第八章 液相色谱电喷雾串联质谱8.1 电喷雾电离源8.2 串联质谱第九章 肽质量指纹谱鉴定蛋白质技术9.1 肽质量指纹谱鉴定蛋白质原理9.2 凝胶电泳分离蛋白质PMF鉴定方法9.3 凝胶电泳分离蛋白质PMF鉴定方法实例第十一章 肽序列标签与蛋白质从头测序技术10.1 串联质谱测定多肽序列原理10.2 串联质谱分析肽段序列原理10.3 肽序列标签鉴定技术10.4 PDS质谱数据库检索鉴定技术10.5 从头测序技术第十一章 植物磷酸化蛋白质的鉴定11.1 磷酸化蛋白质分析概述11.2 磷酸化蛋白质的检测11.3 蛋白质磷酸化位点的分析

5、11.4 蛋白质磷酸化的定量分析第十二章 植物糖基化蛋白质的鉴定12.1 糖蛋白的结构特征12.2 生物质谱技术鉴定糖蛋白12.3 糖蛋白与蛋白质组学第十三章 利用荧光染料进行定量的植物蛋白质组分析技术13.1 二维电泳凝胶的荧光染色13.2 蛋白质样品的荧光标记与分离13.3 图像分析与数据定量第十四章 利用荧光染料进行定量的植物蛋白质组分析技术14.1 运用质谱进行定量的植物蛋白质组分析技术14.2 稳定同位素代谢标记技术14.3 同位素亲和标签技术第十五章 植物蛋白质组研究中的生物信息学15.1 生物信息学简介15.2 数据库的构建15.3 植物蛋白质组研究中常用的网站及数据库全英文教材

6、： Plant Proteomics, Methods and Protocols. in：METHODS IN MOLECULAR BIOLOGY, 355, Edited by Herv Thiellement; Michel Zivy; Catherine Damerval; Valrie Mchin. Human Press,2007.主要参考书：1. Proteomics: From protein sequence to function. Eds S. Pennington & M. Dunn. 2001. BIOS Scientific Publishers2蛋白质组学：理论与

7、方法，钱小红、贺福初 主编，2003年6月，科学出版社出版。3. 国内外相关期刊杂志。Syllabus of Graduate Courses in English in College of Life Science and TechnologyCourse Name: Plant ProteomicsCourse Code:Course Type: For Doctoral Students For Master StudentsAssessment: Examination in English Teaching Language: EnglishFor Majors: Botany,

8、Biochemistry & Molecular Biology, Crop Genetic Breeding Level： Master DoctoralSemester: Fall Credit Hours：Credits: 2Prerequisite Courses: Biochemistry, Molecular BiologyTeachersTitleMajorAgeResearch InterestsGuangxiao YangProf.Biochem. & Mol BiolPlant Biochem. & Mol. Biol.Zhiming TuAssociate Prof.Ov

9、ersea Experience and Academic Expertise of the Course Principal Instructor:1996.101999.03: University of Tsukuba, Japan Dept. of Biological Science Postdoctoral fellow1999.042005.03: National Institute of Agrobiological Resource, Japan, Dept. Of Molecular Genetics, Research ScientistResearch interes

10、ts: Understanding how plants adapt to environmental stresses such as drought, salinity, high and low temperature that result in heavy crop yield losses. We are using genomics, proteomics, biochemical, physiological and bioinformatics approaches to elucidate the mechanisms involved in abiotic stress

11、tolerance in cereals such as wheat and rice. Research interests also include phytohormone signaling and regulated gene expression, the relationships between the structure of storage protein and wheat milling bread making quality.Course Objectives: Proteomics, born in the middle of nineties, twentiet

12、h century, is a new emerging discipline and the frontier of life science research. Proteomics is defined as to study proteome and protein-protein interaction with application of large-scale protein separation and identification techniques. This course focuses on the basic principles and methods in p

13、lant proteomics, and at the same time, introduces the latest advances in the field of plant proteomics. It aims to make students understand and master the research method and experimental technologies including large scale protein separation, identification and protein-protein interactions.Course Ou

14、tline:Chapter 1 Introduction 1.1 Concept in proteomics 1.2 Relation and difference between proteomics and genomics 1.3 Significance and contents of plant proteomics research1.4 Current situation and development tendency in plant proteomics research Chapter 2 Methodology in Proteomics Research2.1 Out

15、line2.2 Large scale protein separation2.3 High-throughput protein identification technologyChapter 3 Protein Extraction from Plant Materials3.1 Outline3.2 Total protein extraction with TCA-Acetone3.3 Extraction of proteins from Recalcitrant plant tissues3.4 Extraction of proteins from woody plant ti

16、ssues3.5 Extraction of proteins from seeds3.6 Extraction of proteins from plant cell organellesChapter 4 Protein Separation Technology4.1 One dimensional isoelectric focusing electrophoresis 4.2 Blue-native gel technique for the analysis of plant protein complexes4.3 Two dimensional SDS - polyacryla

17、mide gel electrophoresis4.4 Protein spot detection on gel.4.5 Truble shootingChapter 5 Image analysis and protein profiling database establishment5.1 Protein electrophoresis image analysis system5.2 Software utility5.3 Two-dimensional electrophoresis of protein spectrum databaseChapter 6 Protein Seq

18、uencing and Protein Identification6.1 Principles6.2 Comparison of advantages and disadvantages of protein sequencer and mass spectrometer6.3 Sample preparation6.4 Analysis and data retrievalChapter 7 Matrix assisted laser desorption ionization time of flight mass spectrometry7.1 MALDI ioniztion sour

19、ce7.2 Linear time of flight mass detector7.3 Reflection time of flight mass detector7.4 Sample preparationChapter 8 Liquid chromatography-electrospray ionization tandem mass spectrometry8.1 The electrospray ionization source8.2 Tandem mass spectrometryChapter 9 Peptide mass fingerprinting for the id

20、entification of protein 9.1 Principle of peptide mass fingerprinting for the identification of proteins 9.2 Gel electrophoresis for protein separation and PMF identification 9.3 Case studyChapter 10 Peptide sequence tag and for de novo protein sequencing 10.1 Principle of tandem mass spectrometry fo

21、r the determination of polypeptide sequence 10.2 Peptide sequence tag identification technology10.3 PDS mass spectrometry database retrieval identification10.4 De novo protein sequencingChapter 11 Plant phosphoprotein identification11.1 Overview on phosphoproteome analysis 11.2 Detection of phosphor

22、ylated proteins11.3 Protein phosphorylation site analysis11.4 Quantitative analysis of protein phosphorylation Chapter 12 Glycoproteins identification in plants12.1 Structural characterization of glycoproteins12.2 Biomass spectrometry identification of glycoprotein12.3 Glycoprotein and proteomicsCha

23、pter 13 Using the fluorescent dye for quantitative plant proteome analysis 13.1 Fluorescent staining for two dimensional gel electrophoresis 13.2 Fluorescence labeling and separation of proteins 13.3 Image analysis and quantitative dataChapter 14 Using mass spectrometry for quantitative plant proteo

24、me analysis14.1 Stable isotope metabolic labeling technique14.2 Isotope coded affinity tag technologyChapter 15 Bioinformatics in plant proteome research 15.1 Introduction to Bioinformatics15.2 Database construction15.3 Web and database commonly used in plant proteome research Textbook Plant Proteomics, Methods and Protocols. in：chin. Human Press, 2007.References：1. Proteomics：Theory & Method. Eds Xiaohong Qian & Fuchu He.2003. Science Press. Beijing （In Chinese）.2Proteomics:3. All related Journals.