Tandem Affinity Purification Tag.ppt课件.ppt

Background Information Rationale for Using TAP/MS Method The Tandem Affinity Purification Tag TAP Purification Strategy TAP Purification Strategy TAP Purification Strategy TAP Purification Strategy … in detail The Experiment The Experiment …cont’d Some Statistics A Higher-Order Map Linked Complexes Results of the Experiment Some Faults of the TAP Method Conclusions Great Summer Reading * Functional organization of the yeast proteome by systematic analysis of protein complexes Anne-Claude Gavin et al. Anne-Claude Gavin, Markus Bosche, Roland Krause, Paola Grandi, Martina Marzioch, Andreas Bauer, Jorg Schultz, Jens M. Rick, Anne-Marie Michon, Cristina-Maria Cruciat, Marita Remor, Christian Hofert, Malgorzata Schelder, Miro Brajenovic, Heins Ruffner, Alejandro Merino, Karin Klein, Manuela Hudak, David Dickson, Tatjana Rudi, Volker Gnau, Angela Bauch, Sonja Bastuck, Bettina Huhse, Christina Leutwein, Marie-Anne Heurtier, Richard R. Copley, Angela Edelmann, Erich Querfurth, Vladimir Rybin, Gerard Drewes, Manfred Raida, Tewis Bouwmesster, Peer Bork, Bertrand Seraphin, Bernhard Kuster, Gitte Neubauer, and Giulio Superti-Furga (Nature 415. 2002) 叭承拯窖默蜘丸赎歇娘筑肖旨汛牛曙待娩帽宰禄长墓惦据轩钨雹扦呸淡姿Tandem Affinity Purification Tag.ppt课件Tandem Affinity Purification Tag.ppt课件 Proteins rarely act alone Comprehensive protein interaction studies thus far: two-hybrid systems (ex vivo) protein chips (in vitro) GST pull-downs (in vivo) 3. Authors used tandem-affinity purification (TAP) and mass spectrometry 匙涸纠粗纺顷肿不铱绘拳窃鼓溢逾焊慈急踏峨东攻楼集跨喂届批奏件旅氏Tandem Affinity Purification Tag.ppt课件Tandem Affinity Purification Tag.ppt课件 Fast purification with high yield In vivo Over-expression of proteins not required Prior knowledge of protein complex not required Purified complex can be used in a variety of studies 蠢视惑骑断葡杂巢蹭慨政跃恿贰桶茧伶宅筷蚁辐答梗彩全甄紫骗懒声酥楚Tandem Affinity Purification Tag.ppt课件Tandem Affinity Purification Tag.ppt课件 target protein target protein 睬绣蜂远哦噪氨冲庶课衬竿蹬耀芯徐翰湛爬吏润磋硬弯湖栈稿赦防请异蓝Tandem Affinity