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The effort to sequence the human genome is now moving toward a c- clusion. As all of the protein coding sequences are described, an increasing emphasis will be placed on understanding gene function and regulation. One important aspect of this analysis is the study of how transcription factors re- late transcriptional initiation by RNA polymerase II, which is responsible for transcribing nuclear genes encoding messenger RNAs. The initiation of Class II transcription is dependent upon transcription factors binding to DNA e- ments that include the core or basal promoter elements, proximal promoter elements, and distal enhancer elements. General initiation factors are involved in positioning RNA polymerase II on the core promoter, but the complex - teraction of these proteins and transcriptional activators binding to DNA e- ments outside the core promoter regulate the rate of transcriptional initiation. This initiation process appears to be a crucial step in the modulation of mRNA levels in response to developmental and environmental signals. Transcription Factor Protocols provides step-by-step procedures for key techniques that have been developed to study DNA sequences and the protein factors that regulate the transcription of protein encoding genes. This volume is aimed at providing researchers in the field with the well-detailed protocols that have been the hallmark of previous volumes of the Methods in Molecular ™ Biology series.
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In Transcription Factor Protocols, Martin Tymms has created a powerful compendium of the major techniques for the study of those DNA sequences and protein factors that regulate the transcription of protein encoding genes. With chapters contributed by leading investigators well versed in the methods, this eminently practical compendium includes not only well established protocols, but also novel techniques that are now being widely adopted. Among the important new methods treated are the use of triplex-forming oligonucleotides, the application of whole genome PCR to the isolation of gene promoters/enhancers, the analysis of in vivo methylation, and in vivo footprinting using UV light and ligation-mediated PCR. Step-by-step instructions, tips about pitfalls to avoid, and extensive procedural notes all help to ensure robust and reproducible results.Transcription Factor Protocols offers both experienced workers and new researchers a vital first-stop reference collection of the core techniques for all those exploring the role of transcription factors in gene regulation today.
Titel: Transcription Factor Protocols
Verlag: Humana
Erscheinungsdatum: 1999
Einband: Hardcover
Zustand: Brand New