Buch, Englisch, Band 545, 345 Seiten, Format (B × H): 198 mm x 266 mm, Gewicht: 1060 g
Reihe: Methods in Molecular Biology
Methods and Protocols
Buch, Englisch, Band 545, 345 Seiten, Format (B × H): 198 mm x 266 mm, Gewicht: 1060 g
Reihe: Methods in Molecular Biology
ISBN: 978-1-60327-992-5
Verlag: Humana Press
in putting togetherthissetofmethods,aswellasallthemembersofmylaboratoryforproofreading and correcting the chapters and especially Sarah McClelland who read all the chapters atleasttwiceatvariousstages. Finally,IwishtothankLyndyRasmusenforherhelpin administeringtheprojectandkeepingmeorganized. Andrew D. McAinsh v Contents Preface. v Contributors. ix 1 Knock-inandKnock-out:TheUseofReverseGeneticsinSomatic CellstoDissectMitoticPathways. 1 AlexisR. Barr, DeborahZyssandFanniGergely 2 FunctionalDissectionofMitoticRegulatorsThroughGene TargetinginHumanSomaticCells. 21 EliBerdougo, Marie-EmilieTerretandPrasadV. Jallepalli 3 RNAiinDrosophilaS2CellsasaToolforStudyingCell CycleProgression. 39 Monica ´ Bettencourt-DiasandGohtaGoshima 4 ProductionofMitoticRegulatorsUsinganAutoselectionSystem forProteinExpressioninBuddingYeast. 63 MarcoGeymonat, AdonisSpanosandStevenSedgwick 5 HydrodynamicAnalysisofHumanKinetochoreComplexes DuringMitosis. 81 SarahE. McClellandandAndrewD. McAinsh 6 IsolationofProteinComplexesInvolvedinMitosisandCytokinesis fromDrosophilaCulturedCells. 99 PierPaoloD’Avino,VincentArchambault,MarcinR. Przewloka, WeiZhang,ErnestD. LaueandDavidM. Glover 7 AutomatedLiveMicroscopytoStudyMitoticGeneFunctionin FluorescentReporterCellLines. 113 MichaelH. A. SchmitzandDanielW. Gerlich 8 ElectronTomographyofMicrotubuleEnd-Morphologies inC. elegansEmbryos. 135 EileenO’TooleandThomasMuller-Reicher ¨ t 9 DissectingMitosiswithLaserMicrosurgeryandRNAi inDrosophilaCells. 145 AntonioJ. Pereira,IrinaMatos,MarianaLince-Faria ´ andHelderMaiato 10 FluorescenceImagingoftheCentrosomeCycleinMammalianCells. 165 SuzannaL. ProsserandAndrewM. Fry vii viii Contents 11 VisualizationofFluorescence-TaggedProteinsinFissionYeast:The AnalysisofMitoticSpindleDynamicsUsingGFP-Tubulin UndertheNativePromoter. 185 MasamitsuSato, MikaToyaandTakashiToda 12 AnalysingKinetochoreFunctioninHumanCells:Spindle CheckpointandChromosomeCongression. 205 ChristianeKlebig,AlbertoToso,SatyarebalaBorusu andPatrickMeraldi 13 ProbingKinetochoreStructureandFunctionUsingXenopuslaevis FrogEggExtracts. 221 MichaelJ. EmanueleandP.
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Weitere Infos & Material
Knock-in and Knock-out: The Use of Reverse Genetics in Somatic Cells to Dissect Mitotic Pathways.- Functional Dissection of Mitotic Regulators Through Gene Targeting in Human Somatic Cells.- RNAi in Drosophila S2 Cells as a Tool for Studying Cell Cycle Progression.- Production of Mitotic Regulators Using an Autoselection System for Protein Expression in Budding Yeast.- Hydrodynamic Analysis of Human Kinetochore Complexes During Mitosis.- Isolation of Protein Complexes Involved in Mitosis and Cytokinesis from Drosophila Cultured Cells.- Automated Live Microscopy to Study Mitotic Gene Function in Fluorescent Reporter Cell Lines.- Electron Tomography of Microtubule End-Morphologies in C. elegans Embryos.- Dissecting Mitosis with Laser Microsurgery and RNAi in Drosophila Cells.- Fluorescence Imaging of the Centrosome Cycle in Mammalian Cells.- Visualization of Fluorescence-Tagged Proteins in Fission Yeast: The Analysis of Mitotic Spindle Dynamics Using GFP-Tubulin Under the Native Promoter.- Analysing Kinetochore Function in Human Cells: Spindle Checkpoint and Chromosome Congression.- Probing Kinetochore Structure and Function Using Xenopus laevis Frog Egg Extracts.- Live Cell Imaging of Kinetochore Capture by Microtubules in Budding Yeast.- The Spindle Checkpoint: Assays for the Analysis of Spindle Checkpoint Arrest and Recovery.- Measuring Proteolysis in Mitosis.- An In Vitro Assay for Cdc20-Dependent Mitotic Anaphase-Promoting Complex Activity from Budding Yeast.- In Vitro Assays for the Anaphase-Promoting Complex/Cyclosome (APC/C) in Xenopus Egg Extracts.- Preparation of Synchronized Human Cell Extracts to Study Ubiquitination and Degradation.- Biochemical Analysis of the Anaphase Promoting Complex: Activities of E2 Enzymes and Substrate Competitive (Pseudosubstrate) Inhibitors.
