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TitleIn Vitro Mutagenesis Protocols [electronic resource] : Third Edition
Author edited by Jeff Braman
ImprintTotowa, NJ : Humana Press : Imprint: Humana Press, 2010
Edition 3
Connect tohttp://dx.doi.org/10.1007/978-1-60761-652-8
Descript XIV, 442 p. 92 illus., 9 illus. in color. online resource

SUMMARY

In the post-genomic era, in vitro mutagenesis has emerged as a critically important tool for establishing the functions of components of the proteome. The third edition of In Vitro Mutagenesis Protocols represents a practical toolbox containing protocols vital to advancing our understanding of the connection between nucleotide sequence and sequence function. Fully updated from the previous editions, this volume contains a variety of specialty tools successfully employed to unravel the intricacies of protein-protein interaction, protein structure-function, protein regulation of biological processes, and protein activity, as well as a novel section on mutagenesis methods for unique microbes as a guide to the generalization of mutagenesis strategies for a host of microbial systems. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and expert tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, In Vitro Mutagenesis Protocols, Third Edition offers today's researchers a valuable compendium of reliable and powerful techniques with which to illuminate the proteome and its rich web of biological implications


CONTENT

Mutagenesis in Various Microbial Backgrounds -- Mutagenesis Protocols in Saccharomyces cerevisiae by In Vivo Overlap Extension -- In Vitro Mutagenesis of Brucella Species -- Random Mutagenesis Strategies for Campylobacter and Helicobacter Species -- Mutagenesis of the Repeat Regions of Herpesviruses Cloned as Bacterial Artificial Chromosomes -- An Efficient Protocol for VZV BAC-Based Mutagenesis -- A Method for Rapid Genetic Integration into Plasmodium falciparum Utilizing Mycobacteriophage Bxb1 Integrase -- PCR Mutagenesis -- Random Mutagenesis by Error-Prone PCR -- A Rapid and Versatile PCR-Based Site-Directed Mutagenesis Protocol for Generation of Mutations Along the Entire Length of a Cloned cDNA -- Rapid Sequence Scanning Mutagenesis Using In Silico Oligo Design and the Megaprimer PCR of Whole Plasmid Method (MegaWHOP) -- Insertion and Deletion Mutagenesis by Overlap Extension PCR -- Targeted Amplification of Mutant Strands for Efficient Site-Directed Mutagenesis and Mutant Screening -- A Modified Inverse PCR Procedure for Insertion, Deletion, or Replacement of a DNA Fragment in a Target Sequence and Its Application in the Ligand Interaction Scan Method for Generation of Ligand-Regulated Proteins -- Amplification of Orthologous Genes Using Degenerate Primers -- Reviews -- Computational Evaluation of Protein Stability Change upon Mutations -- Approaches for Using Animal Models to Identify Loci That Genetically Interact with Human Disease-Causing Point Mutations -- Protein Evolution Mutagenesis -- Using Peptide Loop Insertion Mutagenesis for the Evolution of Proteins -- Massive Mutagenesis®: High-Throughput Combinatorial Site-Directed Mutagenesis -- Directed In Vitro Evolution of Reporter Genes Based on Semi-Rational Design and High-Throughput Screening -- Ribosome Display for Rapid Protein Evolution by Consecutive Rounds of Mutation and Selection -- Protein Structure and Function Mutagenesis -- Fine-Tuning Enzyme Activity Through Saturation Mutagenesis -- Characterization of Structural Determinants of Type 1 Corticotropin Releasing Hormone (CRH) Receptor Signalling Properties -- Site-Directed Mutagenesis for Improving Biophysical Properties of VH Domains -- Phenotype Based Functional Gene Screening Using Retrovirus-Mediated Gene Trapping in Quasi-Haploid RAW 264.7 Cells -- Site-Directed Disulfide Cross-Linking to Probe Conformational Changes of a Transporter During Its Functional Cycle: Escherichia coli AcrB Multidrug Exporter as an Example -- Site-Specific Incorporation of Extra Components into RNA by Transcription Using Unnatural Base Pair Systems -- Random Mutagenesis -- Mutagen™: A Random Mutagenesis Method Providing a Complementary Diversity Generated by Human Error-Prone DNA Polymerases -- Random-Scanning Mutagenesis -- Easy Two-Step Method for Randomizing and Cloning Gene Fragments -- Mutator Bacterial Strain Mutagenesis -- Random Mutagenesis Using a Mutator Strain -- En Passant Mutagenesis: A Two Step Markerless Red Recombination System


Chemistry Human genetics Genetic engineering Proteomics Chemistry Genetic Engineering Human Genetics Proteomics



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