Overview

The pivotal role played by transcription factors in regulating gene expression has given rise to much research and progress in this area. This volume provides detailed practical guidance for identifying, isolating, and characterizing transcription factors. Starting with a specific DNA sequence, these protocols allow the detection and identification of proteins that bind to it and detailed analysis of the DNA binding site. This is followed by procedures for the functional assay of transcription factors and their purification. Also included are methods for the characterization of transcription factor genes, including DNA cloning. The analysis of transcription factors is of central interest to molecular biologists studying gene expression, and this text should be of use to post-graduate students and established researchers alike.

Full Product Details

Author:   David S. Latchman
Publisher:   Oxford University Press
Imprint:   Oxford University Press
Volume:   No. 120
Dimensions:   Width: 15.00cm , Height: 1.10cm , Length: 23.00cm
Weight:   0.390kg
ISBN:  

9780199633418

ISBN 10:   019963341
Pages:   223
Publication Date:   01 March 1993
Audience:   College/higher education ,  Professional and scholarly ,  Postgraduate, Research & Scholarly ,  Professional & Vocational
Replaced By:   9780199636969
Format:   Paperback
Publisher's Status:   Active
Availability:   To order  
Stock availability from the supplier is unknown. We will order it for you and ship this item to you once it is received by us.

Table of Contents

"Part 1 The DNA mobility shift assay, C.L. Dent and D.S. Latchman: detection of DNA-binding proteins; other sources of protein for use in the binding assay; investigation of DNA-binding specificity; characterization of DNA-binding proteins; study of protein-protein interactions. Part 2 Determination of DNA sequences that bind transcription factors by DNA footprinting, N.D. Lakin: preparation of DNA fragments labelled on one strand of the DNA duplex; determination of a transcription factor binding site by DNase I footprinting; determination of a transcription factor binding site by dimethyl sulphate protection footprinting; determination of a transcription factor binding site by dimethyl sulphate interference footprinting; ""in vivo"" footprinting by ligation-mediated polymerase chain reaction. Part 3 Biochemical characterization of transcription factors, Austin J. Cooney et al: determination of molecular weight of the native factor; determination of the molecular weight of the denatured factor; analysis of the monomer/dimer structure of the DNA-binding forms of a transcription factor; identification and initial characterization of non-DNA-binding transcription factors. Part 4 Purification and cloning of transcription factors, R.H. Nichols and G.H. Goodwin: buffers and solutions; preparation of nuclear extract; DNA-cellulose chromatography; reverse-phase chromatography; production of isolated peptides; design of oligonuleotides for cDNA isolation. Part 5 Cloning transcription factors from cDNA expression library, Ian G. Colwell and Helen C. Hurst: handling bacteriophage expression libraries; screening methods; proving the identity factor. Part 6 Cloning transcription factors by homology, Alan Ashworth: methods for cloning related transcription factors; some examples of the cloning of transcription factors by homology. (part contents)"

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