Caenorhabditis Elegans

Caenorhabditis Elegans

Molecular Genetics and Development

2nd Edition - November 23, 2011
  • Editors: Joel Rothman, Andrew Singson
  • eBook ISBN: 9780080925189
  • Hardcover ISBN: 9780125441728

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Description

An updated edition of the classic Methods in Cell Biology volume 48, this book emphasizes diverse methods and technologies needed to investigate C. elegans, both as an integrated organism and as a model system for research inquiries in cell, developmental, and molecular biology, as well as in genetics and pharmacology. By directing its audience to tried-and-true and cutting-edge recipes for research, this comprehensive collection is intended to guide investigators of C. elegans for years to come.

Key Features

  • Diverse, up-to-date techniques covered will be useful to the broadening community of C. elegans researchers for years to come
  • Chapters written by leaders in the field
  • Tried and true methods deliver busy researchers a one-stop compendium of essential protocols

Readership

Researchers and students in cell, molecular and developmental biology

Table of Contents

  • Contributors

    Preface

    Acknowledgments

    PART I Genetics and Functional Genomics

    Chapter 1: Mapping Mutations in C. elegans

    I. Introduction

    II. Mutant Origination

    III. Mutations That Can Be Propagated as Homozygotes

    IV. Mutations That Cannot Be Propagated as Homozygotes

    V. Modifier Mutations

    VI. SNP Mapping

    VII. WGS and Beyond

    Chapter 2: Specialized Chromosomes and Their Uses in Caenorhabditis elegans

    I. Introduction

    II. Chromosomal Rearrangements

    III. Characterization of Chromosomal Rearrangements

    IV. Engineered Constructs: Transgenic Arrays

    Chapter 3: Genome Engineering by Transgene-Instructed Gene Conversion in C. elegans

    I. Introduction

    II. General Features of the MosTIC Technique and Experimental Outline

    III. Methods

    IV. Materials

    V. Discussion: MosTIC Applications and Derivatives

    VI. Summary

    Chapter 4: RNAi Methods and Screening: RNAi Based High-Throughput Genetic Interaction Screening

    I. Introduction: Large-Scale RNAi Screening in C. elegans

    II. Methods

    III. Materials

    PART II Molecular Biology and Biochemistry

    Chapter 5: Dissection of Genetic Pathways in C. elegans

    I. Introduction

    II. Strategies for Identifying Components in Pathways of Interest

    III. Ordering Genes into Pathways

    IV. Future Outlook

    Chapter 6: Transgenesis in C. elegans

    I. Introduction

    II. Uses for Transgenes in C. elegans

    III. Construction of Transgenes

    IV. Obtaining Transgenic Animals

    V. Perspectives: What Lies on the Horizon?

    VI. Summary

    Chapter 7: RNA Processing in C. elegans

    I. Introduction

    II. RNA Processing in Other Organisms

    III. RNA Processing in C. elegans

    Chapter 8: Analysis of microRNA Expression and Function

    I. Introduction

    II. Rationale, Methods, and Materials

    III. Summary

    Chapter 9: In situ Hybridization of Embryos with Antisense RNA Probes

    I. Introduction

    II. Rationale

    III. Methods

    IV. Animal Preparation and Fixation

    V. Hybridization and Signal Development

    VI. Materials

    VII. Notes

    VIII. Troubleshooting Guide

    IX. Imaging of Stained Embryos

    X. Summary

    Chapter 10: Gene-Centered Regulatory Network Mapping

    I. Introduction

    II. Gene Regulatory Networks

    III. Identifying Gene Regulatory Network Nodes

    IV. Gene Regulatory Network Visualization and Analysis

    V. Future Challenges

    Chapter 11: Affinity Purification of Protein Complexes in C. elegans

    I. Introduction

    II. Generating Tools for Biochemistry

    III. The Optimal Starting Material for Protein Purification

    IV. Isolation of Nuclei from Embryos

    V. Preparing Worm and Embryonic Extract

    VI. Single-step Immunoprecipitation

    VII. Tandem Affinity Purification Using a LAP Tag

    VIII. Mass Spectrometry & Prioritization for Follow-up Experiments

    IX. Summary

    X. Solutions and Media

    XI. Equipment

    PART III Development

    Chapter 12: Cell Identification and Cell Lineage Analysis

    I. Introduction

    II. Rationale

    III. Resources

    IV. Nomenclature and Conventions

    V. Cell identification and Lineage Analysis

    VI. Materials, Methods, and Protocols

    VII. Discussion

    Chapter 13: The Genetics and Cell Biology of Fertilization

    I. General Introduction/Background

    II. Finding Sterile Mutants

    III. Maintaining Sterile Mutants

    IV. Fecundity Analysis

    V. Ovulation/Ovulation Rates

    VI. Spermatogenesis

    VII. Sperm Migratory Behavior/Tracking

    VIII. Sperm Competition

    IX. Oogenesis and Oocyte Maturation

    X. Assessing Fertilization and Egg Activation in Egg-Sterile Mutants

    XI. Eggshell Production

    XII. Analysis of Fertilization-Specific Gene Products

    XIII. Future Prospects/Issues

    Chapter 14: Imaging Embryonic Morphogenesis in C. elegans

    I. Introduction

    II. Methods

    III. Summary

    Chapter 15: Molecular and Genetic Approaches for the Analysis of C. elegans Neuronal Development

    I. Introduction

    II. Strategies for In Vivo Labeling of Neurons and Compartments

    III. Other Methods for Examining the Development of Neurons

    IV. Functional Dissection of Signaling Pathways Controlling Neuronal Development

    V. Examples of Neuronal Development

    VI. Outlook

    Chapter 16: Methods for Evaluating the Caenorhabditis elegans Dauer State: Standard Dauer-Formation Assay Using Synthetic Daumones and Proteomic Analysis of O-GlcNAc Modifications

    I. Introduction

    II. Rationale

    III. Methods

    IV. Materials

    V. Discussion

    VI. Summary

    Appendix: Major Caenorhabditis elegans Web Resources

    Index

    Volumes in Series

Product details

  • No. of pages: 504
  • Language: English
  • Copyright: © Academic Press 2011
  • Published: November 23, 2011
  • Imprint: Academic Press
  • eBook ISBN: 9780080925189
  • Hardcover ISBN: 9780125441728

About the Serial Editors

Joel Rothman

Andrew Singson