Publisher's Synopsis
Since the publication of the first edition five years ago, a wide range of new methodologies have been developed to facilitate studies on both isolated parts of the genome and the genome as a whole. This new edition has been updated and expanded so that it provides a comprehensive guide to the methods currently available to characterize the function and activity of an individual transcription factor. All the original chapters have been fully updated or rewritten and additional chapters cover the use of in vitro transcription assays, analysis of chromatin structure, use of the genomic binding site assay and analysis of transcription factor modifications. As with the previous edition, the book starts with a series of chapters concerned with characterizing the proteins binding to a specific DNA sequence and then a chapter on more detailed characterization of the protein itself. The next two chapters describe the isolation of cDNA clones encoding a transcription factor using oligonucleotides predicted from protein sequence and screening of a cDNA expression library. Chapter 6 deals with identification of transcription factors based on sequence homology analysis by both experimental screening and database searches. Chapter 7 is a new chapter that describes methods of identifying the target genes of a previously uncharacterized factor. The next chapters deal with analysis of transcription factor function. Chapter 8 deals with general techniques, and then the following chapters cover the specialized techniques of in vitro transcription assays using transcriptionally active nuclear extracts derived from rat brain, and analysis of the effect of transcription factors on chromatin structure. The final chapter describes methods for detecting the phosphorylation and glycosylation state of transcription factors.
