Xenopus Protocols
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Xenopus Protocols : Cell Biology and Signal Transduction

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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.
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Product details

  • Hardback | 489 pages
  • 158 x 232 x 28mm | 961.62g
  • Totowa, NJ, United States
  • English
  • 2006 ed.
  • XVI, 489 p.
  • 1588293629
  • 9781588293626

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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(TM) 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.
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Table of contents

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. Johne Liu

Xenopus tropicalis Oocytes: More Than Just a Beautiful Genome
Jean-Francois 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
Daniele Roche, Genevieve 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 Pique, Jose Manuel Lopez, and Raul Mendez

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 Labbe, 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 Pante

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
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