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Main description:
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 (TM) Biology series.
Contents:
Isolation of Target Gene Promoter/Enhancer Sequences by Whole Genome PCR Method
Dennis K. Watson, Richard Kitching, Calvin Vary, Ismail Kola, and Arun Seth
In Vivo Footprinting Using UV Light and Ligation-Mediated PCR
Gerd P. Pfeifer and Stella Tommasi
Identification of DNaseI Hypersensitive Sites Within Nuclei
Peter N. Cockerill
Analysis of In Vivo Methylation
Hans-Henrik M. Dahl and Wendy M. Hutchison
Detection of Transcription Factor Partners with a Yeast One Hybrid Screen
Michael Sieweke
Inverse PCR (IPCR) for Obtaining Promoter Sequence
Tony Triglia
PCR-Directed Linker Scanning Mutagenesis
Kurt Gustin and Robert D. Burk
Transfection Technologies
Elaine T. Schenborn
The Use of Particle-Mediated Gene Transfer for the Study of Promoter Activity in Somatic Tissues
Charles M. Nicolet and Ning-Sun Yang
Optimizing Electroporation Conditions for the Transformation of Mammalian Cells
William C. Heiser
Calcium Phosphate Transfection of Mammalian Cultured Cells
Elaine T. Schenborn and Virginia Goiffon
DEAE-Dextran Transfection of Mammalian Cultured Cells
Elaine T. Schenborn and Virginia Goiffon
Liposome-Mediated Transfection of Mammalian Cells
Elaine T. Schenborn and Jennifer Oler
Assays for Transcriptional Activity Based on the Luciferase Reporter Gene
S. Roy Himes and M. Frances Shannon
Transient Transfection of Schneider Cells in the Study of Transcription Factors
Guntram Suske
Triplex-Forming Oligonucleotides and Their Use in the Analysis of Gene Transcription
Marina Kochetkova and M. Frances Shannon
Expression and Purification of Histidine-Tagged Transcription Factors
Donna R. Cohen
Generation of Transcription Factors in Rabbit Reticulocyte Lysate Depleted of Endogenous DNA-Binding Protein
Ulrich Kruse, Thorsten T. Ebel, and Albrecht E. Sippel
Electrophoretic Mobility Shift Assays
Peter L. Molloy
In Vitro Promoter Analysis Using Nuclear Extracts and G-Free Cassette Vectors
Martin J. Tymms
In Vitro Transcription Using Competitor Oligonucleotides to Deplete Specific Transcription Factors
Fujiko Watt
Computer Software of Eukaryotic Promoter Analysis
Dan S. Prestridge
Index
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 316
Weight: 464g
Availability: Available
Subcategories: Genetics
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