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Informationen zum Autor Stephen M. King is Professor of Molecular Biology and Biophysics at the University of Connecticut School of Medicine and is also director of the electron microscopy facility. He has studied the structure, function and regulation of dyneins for over 30 years using a broad array of methodologies including classical/molecular genetics, protein biochemistry, NMR structural biology and molecular modeling, combined with cell biological approaches, imaging and physiological measurements. Klappentext Along with its companion volume on intraflagellar transport, this book provides researchers with a comprehensive and up-to-date source of methods for the analysis cilia and flagella, focusing primarily on approaches that have been devised or significantly extended since the last volume of Methods in Cell Biology on this topic (volume 47, 1995). Edited by Stephen M. King and Gregory J. Pazour, the newest installment of this highly acclaimed serial will serve as an essential addition to the study of cilia and flagella. * Covers protocols for cilia and flagella across systems and species * Both classic and state-of-the-art methods readily adaptable across model systems, and designed to last the test of time * Relevant to clinicians interested in respiratory disease, male infertility, and other syndromes, who need to learn biochemical, molecular, and genetic approaches to studying cilia, flagella, and related structures Zusammenfassung A comprehensive source of methods for the analysis cilia and flagella. It covers protocols for cilia and flagella across systems and species. It is suitable for clinicians interested in respiratory disease! male infertility and other syndromes! who need to learn biochemical and genetic approaches to studying cilia! flagella and related structures. Inhaltsverzeichnis 1. RNAi approaches to axonemal motor function in trypanosomes (Hill) 2. Homologous recombination to knock out dynein genes in Tetrahymena (Gaertig) 3. RNAi in Chlamydomonas (Cerutti) 4. Planarians as a model system for analysis of ciliary assembly/motility (Rompolas) 5. Role of nucleotides in dynein function (Shingyoji) 6. Protein modification to probe dynein interactions + purification and identification of crosslinked dynein products (Sakato) 7. Analysis of redox-sensitive dynein components (Wakabayashi) 8. Purification of dyneins from Chlamydomonas and analysis of dynein-dynein linkers (Kamiya/King) 9. Kinase/phosphatase-mediated control of dynein function (Sale) 10. Central pair MT complex and associated kinesins (Mitchell) 11. Calcium regulation of axonemal function In vitro motility assays (Smith) 12. Identification and analysis of dynein regulatory complex components (Porter) 13. Isolation and analysis of radial spoke proteins (Yang) 14. Purification of dyneins from other model organisms: Ciona, sea urchin, fish (Inaba) 15. Rescue of mutant phenotypes by protein electroporation (Kamiya Lab) 16. Tubulin/Dynein interactions (Raff) 17. Measurement of beat frequency and swimming velocity Cell model reactivation in vitro assays of dynein function (Kamiya) 18. Live imaging of ependymal cilia (Lechtreck/Witman) 19. Immunogold labeling of axonemal components in situ + flat embedding (Geimer) 20. CryoEM approaches to axonemal organization (Nicastro) 21. CryoEM of dynein-microtubule complexes (Oda/Kikkawa) 22. Bioinformatic approaches to dynein HC classification (Yagi) 23. Bioinformatics of non-motor dynein components (Asai) 24. Biophysical approaches to dynein mechanism (Sutoh) 25. Chlamydomonas flagellar beat analysis (Foster) 26. Biophysical measurements of motor function (step size/force production) SAXS of axonemes (Toba/Oiwa) Cryo-Electron Microscope Tomography to Study Axonemal Organization Daniela Nicastro 2. Electron Microscopic Imaging and Analysis of Isolated Dynein Particles Anthony J. Roberts and Stan A. Burgess ...
