Cytoskeleton methods and protocols / edited by Ray H. Gavin.

Format:

Book

Contributor:

Gavin, Ray H.

Series:

Methods in molecular biology (Clifton, N.J.) ; v.161.

Methods in molecular biology ; v. 161

Language:

English

Subjects (All):

Cytoskeleton--Laboratory manuals.

Cytoskeleton.

Cytoskeletal proteins--Laboratory manuals.

Cytoskeletal proteins.

Cytoskeletal Proteins.

Medical Subjects:

Cytoskeleton.

Cytoskeletal Proteins.

Genre:

Laboratory manuals.

Physical Description:

xiv, 288 pages : illustrations (some color) ; 24 cm.

Place of Publication:

Totowa, N.J. : Humana Press, 2001.

Summary:

Ray H. Gavin brings together an international panel of experienced researchers to detail the readily reproducible methods that utilize biochemistry, immunology, genetics, microscopy, and image analysis for investigating cytoskeleton structure and function. Each protocol contains proven step-by-step instructions that enable both the novice and the experienced researcher to achieve successful experimental results. The protocols utilize diverse model systems in a variety of organisms, including Saccharomyces, Micrasterias, Tetrahymena, Drosophila, Spisula, and Xenopus. Microscopy applications include digital-video microscopy and computer-assisted systems for the evaluation of cell motility and morphology. State-of-the-art and highly practical, Cytoskeleton Methods and Protocols makes available a diverse collection of powerful experimental systems and tools for successfully studying cytoskeleton structure and function.

Contents:

Part I. Identification of Cytoskeleton Proteins

1 Using an Inverse PCR Strategy to Clone Large, Contiguous Genomic DNA Fragments / Jorge A. Garces, Ray H. Gavin 3

2 Microsequencing of Myosins for PCR Primer Design / Elaine L. Bearer 9

3 Evaluating the Dynein Heavy Chain Gene Family in Tetrahymena / Gangadhara Sailaja, Leslie M. Lincoln, Jifan Chen, David J. Asai 17

Part II. Microscopy Applications

4 Video-Enhanced Microscopy for Analysis of Cytoskeleton Structure and Function / George M. Langford 31

5 Computer-Assisted Systems for the Analysis of Amoeboid Cell Motility / David R. Soll, Deborah Wessels, Edward Voss, Olof Johnson 45

6 Evaluation of Individual-Cell Motility / Peter S. Walmod, Rasmus Hartmann-Petersen, Anton Berezin, Soren Prag, Vladislav V. Kiselyov, Vladimir Berezin, Elisabeth Bock 59

7 Evaluation of Cell Morphology / Eugene A. Lepekhin, Peter S. Walmod, Anton Berezin, Vladimir Berezin, Elisabeth Bock 85

8 A Quantitative Assay for Measurement of Chemokinesis in Tetrahymena / Uffe Koppelhus, Per Hellung-Larsen, Vagn Leick 101

Part III. Reagents for Studying Cytoskeleton Protein Function

9 Jasplakinolide: An Actin-Specific Reagent that Promotes Actin Polymerization / Andreas Holzinger 109

10 Site-Directed Antibodies as Tools For Investigating Structure and Function of Cytoskeleton Proteins / Walter Steffen, Julie L. Hodgkinson 121

11 Direct Labeling of Components in Protein Complexes by Immuno-Electron Microscopy / Julie L. Hodgkinson, Walter Steffen 133

12 Tat-Mediated Delivery of Antibodies into Cultured Cells / Walter Steffen 141

Part IV. Cytoskeleton Dynamics

13 Studying Cytoskeletal Dynamics in Living Cells Using Green Fluorescent Protein / Yisang Yoon, Kelly R. Pitts, Mark A. McNiven 151

14 Use of Green Fluorescent Protein to Study Cellular Dynamics: Constructing GFP-Tagged Motor Enzymes / Hong Cao, Heather M. Thompson, Eugene W. Krueger, Mark A. McNiven 165

15 Transient Transfections and Heterokaryons as Tools for the Analysis of Keratin IF Dynamics / Jesus M. Paramio, Jose L. Jorcano 189

Part V. Cellular Systems as Tools for Investigating Cytoskeleton Structure and Function

16 Chromatophores as Tools for the Study of Organelle Transport / Bruce R. Telzer, Leah T. Haimo 201

17 Centriole Duplication, Centrosome Maturation, and Spindle Assembly in Lysates of Spisula solidissima Oocytes / Andrew W. Suddith, Eugeni A. Vaisberg, Sergei A. Kuznetsov, Walter Steffen, Conly L. Rieder, Robert E. Palazzo 215

18 Xenopus Egg Extracts as a Model System for Analysis of Microtubule, Actin Filament, and Intermediate Filament Interactions / Craig A. Mandato, Kari L. Weber, Anna J. Zandy, Thomas J. Keating, William M. Bement 229

19 Detergent-Extracted Models for the Study of Cilia or Flagella / Charles B. Lindemann, Kathleen A. Schmitz 241

Part VI. Genetic Approaches for Studies of Cytoskeleton Protein Function

20 A Yeast Two-Hybrid Approach for Probing Cytoskeletal Protein Interactions / Jin-jun Meng, Wallace Ip 255

21 Manipulating Dynein Genes in Tetrahymena / David J. Asai, K. Mark DeWall, Leslie M. Lincoln, Renotta K. Smith 269

22 Functional Analysis of Cytoskeletal Components in the Developing Visual System of Drosophila / Qi He 279.

Notes:

Includes bibliographical references and index.

ISBN:

0896037711

OCLC:

43684635