Drosophila: Methods and Protocols

ISBN-13: 9781588298171 / Angielski / Twarda / 2008 / 452 str.

Dahmann, Christian

Opis

Drosophila melanogaster has been the model system of choice for many inv- tigators over the past hundred years. Due to its long-standing tradition as a model organism, many techniques used in Drosophila have been established and c- tinue to be developed to answer a variety of scientific questions. The recent inv- tion of techniques allowing the knock-down of genes by RNA interference and gene replacement by homologous recombination are two important demonst- tions of the ongoing efforts to broaden the technical repertoire at hand for researchers working with Drosophila. This volume is a collection of protocols covering both standard techniques as well as novel methods. An introductory chapter highlights the importance of Drosophila as a model system for the dev- opment of Biology in the 20th century and review chapters provide concise and up-to-date overviews on selected experimental systems. This book makes no attempt to be comprehensive. A number of frequently used standard techniques were selected and are described in-depth to allow novices to get started with Drosophila. In addition, detailed protocols of recently developed methods that we felt will become of broad use in the Drosophila community within the next years are included. I am grateful to Konrad Basler, Suzanne Eaton, Elisabeth Knust, Pavel Tomancak, and Andreas Wodarz for alerting me to novel techniques and for their advice on selecting the protocols for this book. I would also like to thank all the authors for their expert contribution to this Volume. Christian Dahmann v Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Spis treści

Methods in Molecular Biology Series Editor: John M. Walker Volume: Drosophila: Methods and Protocols Edited by Christian Dahmann, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany Contents Preface Contributors 1 Drosophila melanogaster and the Development of Biology in the XX century Alfonso Martinez Arias 2 Getting started: An overview on raising and using Drosophila Hugo Stocker and Peter Gallant 3 FlyBase: A database for the Drosophila research community Rachel Drysdale 4 The use of P-element transposons to generate transgenic flies André Bachmann and Elisabeth Knust 5 The GAL4 system: a versatile system for the expression of genes David A. Elliott and Andrea H. Brand 6 P-element mutagenesis Thomas Hummel and Christian Klämbt 7 EMS Screens: From mutagenesis to screening and mapping Christian Bökel 8 RNAi screening in cultured Drosophila cells Sandra Steinbrink and Michael Boutros 9 Methods for homologous recombination in Drosophila Keith A. Maggert, Wei J Gong, and Kent G. Golic 10 Recombinases and their use in gene activation, gene inactivation, and transgenesis Johannes Bischof and Konrad Basler 11 Cuticle preparation of Drosophila embryos and larvae Cyrille Alexandre 12 Immunolabelling of embryos H.- Arno J. Müller 13 Imaging cellular and molecular dynamics in live embryos using fluorescent proteins Matthieu Cavey and Thomas Lecuit 14 Analysis of neuromuscular junctions: histology and in vivo imaging Andreas Schmid and Stephan J. Sigrist 15 Immunolabelling of imaginal discs Thomas Klein 16 Imaging Drosophila pupal wing morphogenesis Anne-Kathrin Classen, Benoit Aigouy, Angela Giangrande, and SuzanneEaton 17 Microscopic analysis of the adult Drosophila retina using semi thin plastic sections Konstantin Gaengel and Marek Mlodzik 18 Fluorescent in situ hybridization protocols in Drosophila embryos and tissues Eric Lécuyer, Neela Parthasarathy, and Henry M. Krause 19 Linear RNA amplification for the production of microarray hybridization probes Ansgar Klebes and Thomas B. Kornberg 20 An overview of the identification, detection, and functional analysis of Drosophila microRNAs Nicholas Sokol 21 Extraction and immunoblotting of proteins from embryos Andreas Wodarz 22 Purification of Drosophila protein complexes for mass spectrometry Christoph Jüschke and Jürgen A. Knoblich 23 Mass production of Drosophila embryos and chromatographic purification of native protein complexes Natascha Kunert and Alexander Brehm 24 Flow cytometric analysis of Drosophila cells Aida Flor A. de la Cruz and Bruce A. Edgar 25 Drosophila cell lines as model systems and as an experimental tool Buzz Baum and Lucy Cherbas Index

Blurb

With its long-standing tradition as a model organism, many techniques using Drosophila melanogaster have been established and continue to be developed, while the recent invention of techniques allowing the knock-down of genes by RNA interference and gene replacement by homologous recombination demonstrates the ongoing efforts to broaden the technical repertoire.  In Drosophila: Methods and Protocols, leading experts contribute a collection of protocols covering standard techniques as well as novel methods. As a volume in the highly successful Methods in Molecular Biology™ series, the chapters contain brief introductions to the subject matter, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and essential, Drosophila: Methods and Protocols allows novices and experts alike to tap into the power of one hundred years of expertise involving this vital model system.