Mobile Genetic Elements

ISBN-13: 9781617373015 / Angielski / Miękka / 2010 / 292 str.

Wolfgang J. Miller; Pierre Capy

Opis

Mobile Genetic Elements: Protocols and Genomic Applications brings together a wide array of transposon-based protocols and stategies for studying genome structure, function, and evolution into a highly practical, single-source volume. Such transposable element (TE)-derived techniques have been applied succe- fully for a variety of purposes ranging from mutagenesis, gene silencing, transgenesis, and their use as polymorphic marker systems. To our knowledge no such synthesis has been presented before. Chapters 2-4 provide a series of DNA hybridization techniques for anal- ing the distribution and dynamics of mobile DNAs at the hosts' genomic level. With the current revolution in genomics and the availability of complete genome sequences, computational analyses provide an extremely powerful tool for i- lating and investigating TEs at the in silico level (Chapter 5). For the analyses of transpositional mechanisms at the biochemical level Chapter 6 provides a detailed protocol for LTR retrotransposons in heterologous host systems. Ch- ters 7-10 are focused on TE-based mutagenesis protocols for studying gene functions in a broad range of organisms. Based on their ubiquitous nature and their activity in creating genomic diversity by integrating novel DNA segments into genomes, TEs provide highly informative sets of polymorphic markers (Chapters 11-13). Finally, the last two chapters are dedicated to their technical applications during transgenesis in arthropods and vertebrates.

Spis treści

Mobile Genetic Elements as Natural Tools for Genome Evolution Wolfgang J. Miller and Pierre Capy Detection of Transposable Elements in Drosophila Salivary Gland Polytene Chromosomes by In Situ Hybridization Christian Biémont, Laurence Monti-Dedieu, and Françoise Lemeunier Fluorescent In Situ Hybridization With Transposable Element Probes to Mitotic Chromosomal Heterochromatin of Drosophila Patrizio Dimitri Southern Blot Analysis of Individual Drosophila Flies Nikolaj Junakovic Computational Analysis of Transposable Element Sequences I. King Jordan and Nathan J. Bowen Biochemical Analysis of Long Terminal Repeat Retrotransposons Andreas Bachmair, Karin Garber, Shin Takeda, Kazuhiko Sugimoto, Tetsuji Kakutani, and Hirohiko Hirochika Tn5 as a Molecular Genetics Tool: In Vitro Transposition and the Coupling of In Vitro Technologies With In Vivo Transposition William S. Reznikoff, Igor Y. Goryshin, and Jerry J. Jendrisak Gene Targeting in Drosophila Gregory B. Gloor TE-Based Mutagenesis Systems in Plants: A Gene Family Approach Michiel Vandenbussche and Tom Gerats The Use of Double-Stranded RNA to Knock Down Specific Gene Activity Mary K. Montgomery The Application of LTR Retrotransposons as Molecular Markers in Plants Alan H. Schulman, Andrew J. Flavell, and T. H. Noel Ellis MITE Display Alexandra M. Casa, Alexander Nagel, and Susan R. Wessler Retroposon Mapping in Molecular Systematics Norihiro Okada, Andrew M. Shedlock, and Masato Nikaido Transformation Systems in Insects David A. O´Brochta and Peter W. Atkinson Transposable Elements for Transgenesis and Insertional Mutagenesis in Vertebrates: A Contemporary Review of Experimental Strategies Zoltán Ivics and Zsuzsanna Izsvák Index

Blurb

Transposable elements (TEs)-DNA sequences that are capable of moving from one chromosome location to another-are found in all living organisms. They have been increasingly investigated in a wide spectrum of species, including bacteria, plants, fungi, and animals. In Mobile Genetic Elements: Protocols and Genomic Applications, leading experts describe in step-by-step detail their most productive transposon-based methods and strategies for studying genome structure, function, and evolution. These readily reproducible techniques cover a broad range, including mutagenesis, transgenesis, gene silencing, and molecular systematics. Among the highlights are a series of DNA hybridization methods for analyzing the distribution and dynamics of mobile DNA at the hosts' genomic level, techniques for studying LTR retrotransposons in heterologous host systems, and mutagenesis protocols for investigating gene functions in a broad range of organisms. Additional methods deal with highly informative sets of polymorphic markers, RNAi technology in gene silencing, and applications during transgenesis. The protocols presented follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
State-of-the-art and highly practical, Mobile Genetic Elements: Protocols and Genomic Applications offers investigators powerful genetic tools for dissecting the function of a specific gene, elaborating on the mechanisms leading to genetic change and diversity, and studying the evolutionary impact of mobile DNA on the biology and evolution of organisms.