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.
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Describes transposon-based methods and strategies for studying genome structure, function, and evolution. This work features reproducible techniques including mutagenesis, transgenesis, gene silencing, and molecular systematics. It also offers investigators genetic tools for dissecting the function of a specific gene.
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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.
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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.
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Includes supplementary material: sn.pub/extras
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Produktdetaljer
ISBN
9781588290076
Publisert
2004-03-22
Utgiver
Vendor
Humana Press Inc.
Høyde
235 mm
Bredde
155 mm
Aldersnivå
Research, UU, P, UP, 05, 06
Språk
Product language
Engelsk
Format
Product format
Innbundet