In Vitro Mutagenesis Protocols -

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Résumé :
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.

Sommaire:
Part I: Mutagenesis in Various Microbial Backgrounds 1. Mutagenesis Protocols in Saccharomyces cerevisiae by in vivo Overlap Extension Miguel Alcalde 2. In vitro Mutagenesis of Brucella Species Thomas A. Ficht, Jianwu Pei, and Melissa Kahl-McDonagh 3. Random Mutagenesis Strategies for Campylobacter and Helicobacter Species Duncan J.H. Gaskin and Arnoud H.M. van Vliet 4. Mutagenesis of the Repeat Regions of Herpesviruses Cloned as Bacterial Artificial Chromosomes (BAC) Yuguang Zhao and Venugopal Nair 5. An Efficient Protocol for VZV BAC-Based Mutagenesis Zhen Zhang, Ying Huang, and Hua Zhu 6. A Method for Rapid Genetic Integration into Plasmodium falciparum Utilizing Mycobacteriophage Bxb1 Integrase Sophie H. Adjalley, Marcus C.S. Lee, and David A. Fidock Part II: PCR Mutagenesis 7. Random Mutagenesis by Error-Prone PCR Elizabeth O. McCullum, Berea A.R. Williams, Jinglei Zhang, and John C. Chaput 8. A Rapid and Versatile PCR-Based Site-Directed Mutagenesis Protocol for Generation of Mutations Along the Entire Length of a Cloned cDNA Vincent Dammai 9. Rapid Sequence Scanning Mutagenesis Using in silico Oligo Design and the Megaprimer PCR of Whole Plasmid Method (MegaWHOP) Ulrich Krauss, Karl-Erich Jaeger, and Thorsten Eggert 10. Insertion and Deletion Mutagenesis by Overlap Extension PCR Jehan Lee, Myeong-Kyun Shin, Dong-Kyun Ryu, Seahee Kim, and Wang-Shick Ryu 11. Targeted Amplification of Mutant Strands for Efficient Site-Directed Mutagenesis and Mutant Screening Lei Young and Qihan Dong 12. 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 Oran Erster and Moti Liscovitch 13. Amplification of Orthologous Genes Using Degenerate Primers Samya Chakravorty and Jim O. Vigoreaux Part III: Reviews 14. Computational Evaluation of Protein Stability Change Upon Mutations Shuangye Yin, Feng Ding, and Nikolay V. Dokholyan 15. Approaches for Using Animal Models to Identify Loci that Genetically Interact with Human Disease-Causing Point Mutations Josef D. Franke Part IV: Protein Evolution Mutagenesis 16. Using Peptide Loop Insertion Mutagenesis for the Evolution of Proteins Christian Heinis and Kai Johnsson 17. Massive Mutagenesis?: High-Throughput Combinatorial Site-Directed Mutagenesis Julien Sylvestre 18. Directed in vitro Evolution of Reporter Genes Based on Semi-Rational Design and High Throughput Screening Ai-Sheng Xiong, Quan-Hong Yao, Ri-He Peng, and Zong-Ming Cheng 19. Ribosome Display for Rapid Protein Evolution by Consecutive Rounds of Mutation and Selection Hayato Yanagida, Tomoaki Matsuura, and Tetsuya Yomo Part V: Protein Structure and Function Mutagenesis 20. Fine-Tuning Enzyme Activity through Saturation Mutagenesis Holly H. Hogrefe 21. Characterization of Structural Determinants of Type 1 Corticotropin Releasing Hormone (CRH) Receptor Signalling Properties Danijela Markovic and Dimitris K. Grammatopoulos 22. Site-Directed Mutagenesis for Improving Biophysical Properties of VH Domains Mehdi Arbabi-Ghahroudi, Roger MacKenzie, and Jamshid Tanha 23. Phenotype Based Functional Gene Screening Using Retrovirus-Mediated Gene Trapping in Quasi-Haploid RAW 264.7 Cells Sung Ouk Kim and Soon-Duck Ha 24. Site-Directed Disulfide Cross-Linking to Probe Conformational Changes of a