蛋白质相互作用-实验方法

实验频道正文用户登录新用户注册蛋白质相互作用作者：佚名实验频道来源：中华基因网点击数：更新时间：2004-6-8 Interaction Trap/Trap Two-Hybrid System · Yeast Two-Hybrid System (Finley Lab)This is one of the most comprehensive and detailed guide to yeast two-hybrid system technique with introduction to its background. The following procedures are described: mating assay - small scale for tens of different bait or prey proteins, collecting bait (and prey) strains, large scale for hundreds of different bait or prey strains, interaction mating assay with other yeast two-hybrid systems, recording results, interpreting interaction data, qualitative interpretation and more...· Yeast Two-Hybrid System (Finley Lab)The updated version for scaling up the these techniques to facilitate characterization of large networks of interacting proteins. It is also useful for individual hunts.· Performing a hunt by interaction mating (Finley Lab) In this protocol one strain is transformed with library DNA and the transformants are collected and frozen in aliquots. For each interactor hunt, an aliquot of this frozen pretransformed library strain is thawed and mixed with an aliquot of a bait strain transformed with the bait expression plasmid. Overnight incubation of the mixture on a YPD plate results in mating - i.e., cells of one strain fuse with cells of the other strain to form diploids. The diploids are then exposed to galactose to induce expression of the library encoded proteins, and interactors are selected in the same manner as in the standard hunt protocols.· Yeast Two-Hybrid System: part A: screen preparation (Technical Tips Online) (To view this page, free registration is needed)The two-hybrid system (THS) is a powerful technique for identifying new proteins involved in specific biological processes. It allows for the rapid isolation of the gene that codes for a protein that interacts with a specific protein of interest. This Protocol describes procedures useful for the `Fields THS, based on the GAL4 transcription factor including o Construction of the GAL4BD:bait gene fusion plasmid o Choosing the right THS reporter strain o Testing the BD:bait plasmid for auto-activation o Expression of your fusion protein o Amplifying the AD:cDNA library · Yeast two-hybrid system: part B--screening procedure (Technical Tips Online) · Yeast two-hybrid system: part C--characterizing positives (Technical Tips Online) · Two Hybrid System Transformation (Gietz Lab)This Protocol is for 2 Hybrid system screensAffinity Purification of GST Fusion ProteinsPurification of GST Fused Proteins (Contributed by Chia Jin Ngee)Many people have vented out frustration over insoluble GST-fused proteins. This is a protocol for enzymatically active soluble GST-fused proteins. All GST-fused proteins are rendered soluble with this technique though enzyme activitiy can range from 30-90%. GST Fusion Protein Purification from Yeast (Dohlman Lab) · Purification of MBP (maLTose-Binding Proteins) Fused Proteins (PMCI) · Purification of His-tagged proteins from Sf9 cells under native conditions (Brent Graveley) Purification of GST Fused Proteins (PMCI) GST-Fusion Protein Purification (Marks Lab) 处理 SSI 文件时出错处理 SSI 文件时出错【评论】【进论坛看】【转入博客】实验频道录入：生物编辑责任编辑：生物编辑上一篇实验频道：蛋白质电泳下一篇实验频道：蛋白质磷酸化【发表评论】【加入收藏】【告诉好友】【打印此文】【关闭窗口】相关文章最新热门推荐文章蛋白质提取与纯化技术2%多聚甲醛固定剂配方二则细胞学技术细胞培养的一般技术血小板的活化、荧光染色与流血小板活化检测方法细胞表面抗原染色的组织样本流式细胞仪PI染色死细胞方法流式细胞术分析血小板流式检测细胞周期 · 免疫细胞表面抗原分子CD家族对照表· 所有的看家基因(housekeeping gene· 生物发光与化学发光专题· 生物发光和化学发光在生物技术中的· 重组DNA的分离、克隆与测序实验手册· RNAi和基因沉默的历史回顾· RNAi原理图解· RNAi术语表· siRNA设计指南· Methods for DNA sequencing · 通用电气医疗集团简介· 生物发光与化学发光专题· 生物发光和化学发光在生物技术中的· RNA干扰技术获得新突破· 蛋白质特性与分离纯化技术的选择· PCR实验指导与常见问题分析· RNAi原理FLASH演示· Extrachromosomal elements, plasm· Promoter analysis by saturation · 重组DNA的分离、克隆与测序实验手册文章评论（只显示最新10条。评论内容只代表网友观点，与本站立场无关！）生物谷 | 网站介绍 | 网站地图 | 广告服务 | 帮助关于我们 | 广告服务 | 战略伙伴 | 留言| 帮助信息| 联系方式 | 客户投诉 | 友情链接 | 网站地图 | 网站声明 | Bioon English 上海北岸信息技术有限公司公司地址：上海龙吴路51号嘉源商务中心1#211室(200232) 生物谷网站 bioon.com © Copyright 2001 - 2006. All rights reserved. 备案号：沪ICP备05022939号
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