Pub. Date:
Elsevier Science
Applications of Chimeric Genes and Hybrid Proteins, Part A: Gene Expression and Protein Purification

Applications of Chimeric Genes and Hybrid Proteins, Part A: Gene Expression and Protein Purification


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Product Details

ISBN-13: 9780121822279
Publisher: Elsevier Science
Publication date: 10/02/2000
Series: Methods in Enzymology Series
Edition description: New Edition
Pages: 617
Product dimensions: 6.30(w) x 9.29(h) x 1.27(d)

Table of Contents

Contributors to Volume 326ix
Volumes in Seriesxv
Section I.Historical Overview
1.The All Purpose Gene Fusion3
Section II.Gene Fusions as Reporters of Gene Expression in Prokaryotic Cells
2.A Practical Guide to the Construction and Use of lac Fusions in Escherichia coli11
3.Tagging Exported Proteins Using Escherichia coli Alkaline Phosphatase Gene Fusions35
4.Applications of Gene Fusions to Green Fluorescent Protein and Flow Cytometry to the Study of Bacterial Gene Expression in Host Cells47
5.IVET and RIVET: Use of Gene Fusions to Identity Bacterial Virulence Factors Specifically Induced in Host Tissues73
6.Identification of Exported Bacterial Proteins via Gene Fusions to Yersinia pseudotuberculosis Invasin97
Section III.Gene Fusions as Reporters of Gene Expression in Eukaryotic Cells
7.Use of Imidazoleglycerolphosphate Dehydratase (His3) as a Biological Reporter in Yeast107
8.Use of Fusions to Human Thymidine Kinase as Reporters of Gene Expression and Protein Stability in Saccharomyces cerevisiae120
9.Use of Fusions to Thymidine Kinase134
10.In Vivo Analysis of lacZ Fusion Genes in Transgenic Drosophila melanogaster146
11.Utility of the Secreted Placental Alkaline Phosphatase Reporter Enzyme159
12.Fusions to Imidazopyrazinone-Type Luciferases and Aequorin as Reporters165
13.Novel Methods for Chemiluminescent Detection of Reporter Enzymes175
14.Fusions to Chloramphenicol Acetyltransferase as a Reporter202
15.Fusions to [beta]-Lactamase as a Reporter for Gene Expression in Live Mammalian Cells221
Section IV.Functional Tags for Biochemical Purification
16.Purification of Proteins Using Polyhistidine Affinity Tags245
17.Generating Fusions to Glutathione S-Transferase for Protein Studies254
18.Use of the Strep-Tag and Streptavidin for Detection and Purification of Recombinant Proteins271
19.Streptavidin-Containing Chimeric Proteins: Design and Production305
20.Fusions to Maltose-Binding Protein: Control of Folding and Solubility in Protein Purification312
21.Thioredoxin as a Fusion Partner for Production of Soluble Recombinant Proteins in Escherichia coli322
22.Affinity Purification of Recombinant Proteins Fused to Calmodulin or to Calmodulin-Binding Peptides340
23.The S Tag Fusion System for Protein Purification362
24.Fusions to Self-Splicing Inteins for Protein Purification376
25.Fusion Proteins Containing Cellulose-Binding Domains418
26.Biotinylation of Proteins in Vivo and in Vitro Using Small Peptide Tags430
27.Biotinylation of Proteins in Vivo: A Useful Post-translational Modification for Protein Analysis440
Section V.Hybrid Proteins for Detection and Production of Antigens and Antibodies
28.Methods for Generating Multivalent and Bispecific Antibody Fragments461
29.Design and Use of Phage Display Libraries for the Selection of Antibodies and Enzymes480
30.Use of an Lpp-OmpA Fusion Vehicle for Bacterial Surface Display506
31.Identification of Bacterial Class I Accessible Proteins by Disseminated Insertion of Class I Epitopes516
32.Bordetella pertussis Adenylate Cyclase Toxin: A Vehicle to Deliver CD8-Positive T-Cell Epitopes into Antigen-Presenting Cells527
33.Use of Anthrax Toxin Fusions to Stimulate Immune Responses542
34.Use of Fusions to Viral Coat Proteins as Antigenic Carriers for Vaccine Development551
Author Index571
Subject Index601

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