9酵母双杂交技术.PPT

HIS3：筛选标记 酵母单杂交技术 Li等（1993）从酵母双杂交技术发展而来，研究DNA和蛋白质互作。 ChIP: Chromatin immuno precipitation 主要实验流程：在活细胞状态下固定蛋白质－DNA复合物，并通过超声或酶处理将其随机切断为一定长度范围内的染色质小片段，然后通过抗原抗体的特异性识别反应沉淀此复合体，特异性地富集目的蛋白结合的DNA片段，通过对目的片段的纯化与检测，获得蛋白质与DNA相互作用的信息。 染色质免疫共沉淀 思考题 什么是基因工程？ 对社会和科学技术发展有什么重要意义？ 限制性内切酶有哪几类？什么是回文结构？常见载体？报告基因？蓝白斑筛选？ 基因克隆的常用策略及原理？果树上使用的有哪些方法？ 什么是细菌感受态？转化？转染？酵母双杂交？ 主要参考资料 基因 VIII 基因操作原理 孙明 华中农业大学 植物基因工程 王关林 方宏均 科学出版社 2002 Molecular Cloning 分子克隆实验指南（E3,2001） Principles of Gene Manipulation (S Primrose, R Twyman, Blackwell scientific publication) Serial analysis of gene expression (SAGE) is a method for comprehensive analysis of gene expression patterns. Three principles underlie the SAGE methodology: 1) A short sequence tag (10-14bp) contains sufficient information to uniquely identify a transcript provided that that the tag is obtained from a unique position within each transcript; 2) Sequence tags can be linked together to from long serial molecules that can be cloned and sequenced; and 3) Quantification of the number of times a particular tag is observed provides the expression level of the corresponding transcript. Serial analysis of gene expression (SAGE) is a method for comprehensive analysis of gene expression patterns. Three principles underlie the SAGE methodology: 1) A short sequence tag (10-14bp) contains sufficient information to uniquely identify a transcript provided that that the tag is obtained from a unique position within each transcript; 2) Sequence tags can be linked together to from long serial molecules that can be cloned and sequenced; and 3) Quantification of the number of times a particular tag is observed provides the expression level of the corresponding transcript. 特定的方法在特定的基质上制造的分子微点阵，利用分子间的特异性相互作用原理，将不连续的分析过程集成于芯片表面。 * The chips are manufactured on wafers. These wafers are 5-inch-by-5-inch pieces of glass. You can get anywhere from 49 to 900 individual chips out of each wafer, depending on the overall size of the final chip to be made A standa