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Identification and Expression of G Protein-Coupled Receptors

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The past decade has seen tremendous advances in the study of G protein-coupled receptors (GPCRs), including the molecular cloning and identification of more than 100 GPCR genes. But while GPCRs serve as targets for more than 300 medicines in modern pharmacology, the shrinking pool of know ligands and the continuing discovery of orphan GPCR genes have underscored the need for new approaches for ligand identification. This book addresses this new direction in GPCR biochemistry, offering a definitive laboratory ...
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Overview

The past decade has seen tremendous advances in the study of G protein-coupled receptors (GPCRs), including the molecular cloning and identification of more than 100 GPCR genes. But while GPCRs serve as targets for more than 300 medicines in modern pharmacology, the shrinking pool of know ligands and the continuing discovery of orphan GPCR genes have underscored the need for new approaches for ligand identification. This book addresses this new direction in GPCR biochemistry, offering a definitive laboratory bench manual that emphasizes expression over primary cloning strategies.

The book contains black-and-white illustrations.

The past decade has seen tremendous advances in the study of G protein-coupled receptors (GPCRs), including the molecular cloning and identification of more than 100 GPCR genes. But while GPCRs serve as targets for more than 300 medicines in modern pharmacology, the shrinking pool of know ligands and the continuing discovery of orphan GPCR genes have underscored the need for new approaches for ligand identification. This book addresses this new direction in GPCR biochemistry, offering a definitive laboratory bench manual that emphasizes expression over primary cloning strategies.

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Editorial Reviews

Eugene A. Davidson
This is a guide to the identification and expression of G protein-coupled receptors. This volume is intended as a ""how-to"" laboratory accessory for investigators wanting to express G protein-coupled receptors. Investigators with a need to express such receptors are the intended audience, but they will have difficulty in decision making since there is a lack of critical commentary on the pros and cons of the various technologies. The coverage is reasonably complete in that the most common methods of protein expression are described—bacteria, mammalian cells, insect cells, oocytes, and yeast. Attention is also given to the growing problem of information management and there is an extensive listing of cloned receptors together with accession data. It is not uncommon for a given protein to be effectively expressed by one method (e.g., yeast) and fail miserably in a different genetic background. Given a new candidate, how does an investigator decide on the best strategic approach? If one is willing to try several, this volume will have value. If resources are limited so that the shotgun approach is not feasible, prudent consultation is likely to provide a better outcome. It also should be noted that several of the chapters provide descriptive material as opposed to protocols. In this regard, the utility of the book as a laboratory adjunct is limited.
Booknews
A laboratory bench manual for biochemists, pharmacologists, neuro- scientists, structural biologists, and other investing the receptors. Emphasizing expression over primary cloning strategies, provides detailed protocols for various expression systems from bacteria to mammalian cells, and offers opinions on the advantages and shortcoming of each approach. Among the topics are homology screening and the polymerase chain reaction in cloning the receptor genes, heterologous expression in primary cell cultures, large-scale expression and purification in mammalian cells, Schneider 2 cells, and methods for genetic analysis and ligand identification using heterologous receptors expressed in . Annotation c. by Book News, Inc., Portland, Or.
Doody's Review Service
Reviewer: Eugene A Davidson, PhD (Georgetown University School of Medicine)
Description: This is a guide to the identification and expression of G protein-coupled receptors.
Purpose: This volume is intended as a "how-to" laboratory accessory for investigators wanting to express G protein-coupled receptors.
Audience: Investigators with a need to express such receptors are the intended audience, but they will have difficulty in decision making since there is a lack of critical commentary on the pros and cons of the various technologies.
Features: The coverage is reasonably complete in that the most common methods of protein expression are described — bacteria, mammalian cells, insect cells, oocytes, and yeast. Attention is also given to the growing problem of information management and there is an extensive listing of cloned receptors together with accession data.
Assessment: It is not uncommon for a given protein to be effectively expressed by one method (e.g., yeast) and fail miserably in a different genetic background. Given a new candidate, how does an investigator decide on the best strategic approach? If one is willing to try several, this volume will have value. If resources are limited so that the shotgun approach is not feasible, prudent consultation is likely to provide a better outcome. It also should be noted that several of the chapters provide descriptive material as opposed to protocols. In this regard, the utility of the book as a laboratory adjunct is limited.

1 Star from Doody
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Product Details

  • ISBN-13: 9780471183105
  • Publisher: Wiley, John & Sons, Incorporated
  • Publication date: 3/5/1998
  • Series: Receptor Biochemistry and Methodology Series , #20
  • Edition number: 1
  • Pages: 222
  • Product dimensions: 7.28 (w) x 10.24 (h) x 0.67 (d)

Table of Contents

Series Preface
Preface
Contributors
1 Cloning of G Protein-Coupled Receptor Genes: The Use of Homology Screening and the Polymerase Chain Reaction 1
2 Practical Approaches for Cloning G Protein-Coupled Receptors Using Mammalian Cell Expression 27
3 G Protein-Coupled Receptor Informatics and the Orphan Problem 54
4 The Use of Xenopus laevis Oocytes for the Study of G Protein-Coupled Receptors 73
5 Stable Expression of G Protein-Coupled Receptors in Mammalian Cells 97
6 Heterologous Expression in Primary Cell Cultures 113
7 Expression of G Protein-Coupled Receptors in Escherichia coli 133
8 Large-Scale Expression/Purification of G Protein-Coupled Receptors: Mammalian Cells 151
9 High-Level Expression of G Protein-Coupled Receptors in the Baculovirus/SF9 Cell Expression System 167
10 Expression of G Protein-Coupled Receptors in Drosophila Schneider 2 Cells 181
11 Heterologous G Protein-Coupled Receptors Expressed in Saccharomyces cerevisiae: Methods for Genetic Analysis and Ligand Identification 196
Index 213
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