In recent years, much information has been revealed concerning the essential role of helicases, the enzymes that utilize the energy derived from nucleosidetriphosphate hydrolysis to unwind the double stranded helical structure of nucleic acids. In Helicases: Methods and Protocols, this topic is explored by leading international scientists who have contributed significantly to our current understanding of helicases and the ways in which they function. Chapters include methods for the production and purification of helicases from different species, as well as detailed studies of helicase activities, including NTP binding and hydrolysis, nucleic acid binding and unwinding, and translocation along nucleic acid substrates. Composed in the highly successful Methods in Molecular Biology™ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Wide-ranging and innovative, Helicases: Methods and Protocols is the ultimate guide for researchers,providing the scientific community with comprehensive accounts of the most advanced methods used to study helicases today. INDICE: Helicases: An overview.- Fluorescent biosensors to investigate helicase activity.- Single molecule FRET analysis of helicase functions.- Kinetics of Motor Protein Translocation on Single Stranded DNA.- Experimental and computational analysis of DNA unwinding and polymerization kinetics.- Protein displacement by helicases.- In vivo Investigation of Protein-Protein Interactionsfor Helicases Using Tandem Affinity Purification.- Mapping genomic targets ofDNA helicases by chromatin immunoprecipitation in Saccharomyces cerevisiae.- Methods to Study How Replication Fork Helicases Unwind DNA.- Simple enzymatic assays for the in vitro motor activity of transcription termination factor Rhofrom Escherichia coli.- Single-molecule studies of RecBCD.- Mutational analysis of Bloom helicase.- In vivo analysis of Drosophila BLM helicase function during DNA double-strand gap repair.- Purification and characterization of RecQ helicases of plants.- Fluorometric assay of hepatitis C virus NS3 helicase activity.- A Method to Simultaneously Monitor Hepatitis C Virus NS3 Helicase and Protease Activities.- Computer Modeling of Helicases Using Elastic Network Model.- Duplex unwinding with DEAD-box proteins.- Analysis of the RNA helicase p68 (Ddx5) as a transcriptional regulator.- A method to study the role of DDX3 RNA helicase in HIV-1.- Molecular characterization of nuclear DNA helicase II (RNA helicase A).- Regulation of inter- and intramolecular interaction of RNA, DNA and proteins by MLE.- Biochemical characterization of human Upf1 helicase.- Assays of the helicase, ATPase and exoribonuclease activities of the yeast mitochondrial degradosome.- Characterization of the Helicase Activity and Anti-Telomerase Properties of Yeast Pif1p in vitro.- A method to confer salinity stress tolerance to plants by helicase overexpression.- A method to inhibit the growth of Plasmodium falciparum by double-stranded RNA-mediated gene silencingof helicases.
- ISBN: 978-1-60327-354-1
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 382
- Fecha Publicación: 01/10/2009
- Nº Volúmenes: 1
- Idioma: Inglés