Full Description
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.
Contents
Mobile Genetic Elements as Natural Tools for Genome Evolution.- Detection of Transposable Elements in Drosophila Salivary Gland Polytene Chromosomes by In Situ Hybridization.- Fluorescent In Situ Hybridization With Transposable Element Probes to Mitotic Chromosomal Heterochromatin of Drosophila.- Southern Blot Analysis of Individual Drosophila Flies.- Computational Analysis of Transposable Element Sequences.- Biochemical Analysis of Long Terminal Repeat Retrotransposons.- Tn5 as a Molecular Genetics Tool.- Gene Targeting in Drosophila.- TE-Based Mutagenesis Systems in Plants.- The Use of Double-Stranded RNA to Knock Down Specific Gene Activity.- The Application of LTR Retrotransposons as Molecular Markers in Plants.- MITE Display.- Retroposon Mapping in Molecular Systematics.- Transformation Systems in Insects.- Transposable Elements for Transgenesis and Insertional Mutagenesis in Vertebrates.
