图书简介
Great disparities exist between organisms with regard to the relative ease of chromosomal mutagenesis and manipulation. In Chromosomal Mutagenesis, a team of experts provide a variety of chromosomal manipulation techniques, including insertional gene disruptions, gene knockouts, stimulated homologous recombination techniques and other novel tools, for both prokaryotic and eukaryotic organisms, and attempt to expand the genetic toolbox beyond model organisms. Following the format of the highly successful Methods in Molecular Biology? format, each chapter offers step-by-step laboratory instructions, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Chromosomal Mutagenesis covers state-of-the-art techniques that are staged to expand, if not revolutionize, genetic analysis in the long neglected and relevant cell types.
Biallelic Gene Knockouts in Chinese Hamster Ovary Cells.- Highly Proficient Gene Targeting by Homologous Recombination in the Human Pre-B Cell Line Nalm-6.- Robust Cell Line Development Using Meganucleases.- Design and Testing of Zinc Finger Nucleases for Use in Mammalian Cells.- Gene Targeting in Drosophila and Caenorhabditis elegans With Zinc-Finger Nucleases.- Orpheus Recombination.- Transposon-Mediated Mutagenesis in Somatic Cells.- Insertional Mutagenesis of the Mouse Germline With Sleeping Beauty Transposition.- Conditional Gene Trapping Using the FLEx System.- Steps Toward Targeted Insertional Mutagenesis With Class II Transposable Elements.- Targeting Integration of the Saccharomyces Ty5 Retrotransposon.- Site-Specific Chromosomal Integration Mediated by ?C31 Integrase.- Triplex-Mediated Gene Modification.- Allelic Exchange of Unmarked Mutations in Mycobacterium Tuberculosis.- Mycobacterial Recombineering.- Chromosomal Engineering of Clostridium Perfringens Using Group II Introns.
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