Xenopus Protocols: Cell Biology and Signal Transduction

ISBN-13: 9781588293626 / Angielski / Twarda / 2005 / 489 str.

X. Johne Lio; X. John Liu; X. Johne Liu

Opis

A collection of standard and cutting-edge techniques for using Xenopus oocytes and oocytes/egg extracts to reconstitute biological and cellular processes. These readily reproducible methods take advantage of the oocyte's impressive protein abundance, its striking protein translation capacity, and its breathtaking possibilities for the assembly of infectious viral particles by single cell injection of multiple RNAs. The authors focus on the versatility of frog oocytes and egg extracts in cell biology and signal transduction, and cover all the major uses of oocytes/extracts as experimental models.

Spis treści

Resources for Genetic and Genomic Studies of Xenopus Steven L. Klein, Daniela S. Gerhard, Lukas Wagner, Paul Richardson, Lynn M. Schriml, Amy K. Sater, Wesley C. Warren, and John D. McPherson The Physiology of the Xenopus laevis Ovary Melissa A. Rasar and Stephen R. Hammes Oocyte Isolation and Enucleation X. Shawn Liu and X. Johné Liu Xenopus tropicalis Oocytes: More Than Just a Beautiful Genome Jean-François L. Bodart and Nicholas S. Duesbery Oocyte Expression With Injection of Purified T7 RNA Polymerase Xavier Altafaj, Nathalie Joux, Michel Ronjat, and Michel De Waard Visualization of the Cytoskeleton in Xenopus Oocytes and Eggs by Confocal Immunofluorescence Microscopy Bret E. Becker and David L. Gard Multiphoton Laser Scanning Microscopy as a Tool for Xenopus Oocyte Research Angela M. Prouty, Jun Wu, Da-Ting Lin, Patricia Camacho, and James D. Lechleiter Imaging Ca2+ Signals in Xenopus Oocytes Sheila L. Dargan, Angelo Demuro, and Ian Parker Chromosomal DNA Replication in a Soluble Cell-Free System Derived From Xenopus Eggs Antonin V. Tutter and Johannes C. Walter Chromatin Assembly of DNA Templates Microinjected Into Xenopus Oocytes Danièle Roche, Geneviève Almouzni, and Jean-Pierre Quivy Pre-mRNA Splicing in the Nuclei of Xenopus Oocytes Kyong Hwa Moon, Xinliang Zhao, and Yi-Tao Yu Chromatin Immunoprecipitation for Studying Transcriptional Regulation in Xenopus Oocytes and Tadpoles David Stewart, Akihiro Tomita, Yun-Bo Shi, and Jiemin Wong Cytoplasmic mRNA Polyadenylation and Translation Assays Maria Piqué, José Manuel López, and Raúl Méndez Xenopus Egg Extracts: A Model System to Study Proprotein Convertases Kathleen I. J. Shennan Analysis of Molecular Chaperones Using a Xenopus Oocyte Protein Refolding Assay John J. Heikkila,Angelo Kaldis, and Rashid Abdulle Ubiquitin-Mediated Protein Degradation in Xenopus Egg Extracts Anna Castro, Suzanne Vigneron, Cyril Bernis, Jean-Claude Labbé, and Thierry Lorca Introduction to Nucleocytoplasmic Transport: Molecules and Mechanisms Reiner Peters Use of Xenopus laevis Oocyte Nuclei and Nuclear Envelopes in Nucleocytoplasmic Transport Studies Reiner Peters Nuclear Pore Complex Structure and Plasticity Revealed by Electron and Atomic Force Microscopy Bohumil Maco, Birthe Fahrenkrog, Ning-Ping Huang, and Ueli Aebi In Vitro Study of Nuclear Assembly and Nuclear Import Using Xenopus Egg Extracts Rene C. Chan and Douglass J. Forbes Use of Intact Xenopus Oocytes in Nucleocytoplasmic Transport Studies Nelly Panté Studying the Mechanosensitivity of Voltage-Gated Channels Using Oocyte Patches Catherine E. Morris, Peter F. Juranka, Wei Lin, Terence J. Morris, and Ulrike Laitko Oocytes as an Expression System for Studying Receptor/Channel Targets of Drugs and Pesticides Steven David Buckingham, Luanda Pym, and David Barry Sattelle Microtransplantation of Neurotransmitter Receptors From Cells to Xenopus Oocyte Membranes: New Procedure for Ion Channel Studies Ricardo Miledi, Eleonora Palma, and Fabrizio Eusebi Reconstitution of Golgi Disassembly by Mitotic Xenopus Egg Extracts in Semi-Intact MDCK Cells Fumi Kano, Katsuya Takenaka, and Masayuki Murata Exploring RNA Virus Replication in Xenopus Oocytes Andrea V. Gamarnik and Raul Andino Study of Apoptosis in Vitro Using the Xenopus Egg Extract Reconstitution System Paula Deming and Sally Kornbluth Studying Fertilization in Cell-Free Extracts: Focusing on Membrane/Lipid Raft Functions and Proteomics Ken-ichi Sato, Ken-ichi Yoshino, Alexander A. Tokmakov, Tetsushi Iwasaki, Kazuyoshi Yonezawa, and Yasuo

Blurb

Although Xenopus oocytes are very hardy and robust in terms of experimentation and offer unusually simple and inexpensive opportunities for the delivery of mRNA to the cytoplasm and DNA to the nucleus, they are underutilized as an experimental model. In Xenopus Protocols: Cell Biology and Signal Transduction, hands-on researchers describe standard and cutting-edge techniques for using Xenopus oocytes and oocyte/egg extracts to reconstitute biological and cellular processes. These readily reproducible methods take advantage of the oocyte's impressive protein abundance (for example, 50-70 ng catalytic subunit of PKA per oocyte), its striking protein translation capacity (200-400 ng protein/oocyte per day), and its breathtaking possibilities for the assembly of infectious viral particles by single-cell injection of multiple RNAs. The authors focus on the versatility of frog oocytes and egg extracts in cell biology and signal transduction and cover all the major uses of oocytes/extracts as experimental models. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and highly practical, Xenopus Protocols: Cell Biology and Signal Transduction serves perfectly to instruct novices in the use of Xenopus oocytes/extracts, as well as seasoned investigators interested in developing novel models and uses.