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A Laboratory Guide to In Vitro Studies of Protein-DNA Interactions (Paperback, Softcover reprint of the original 1st ed. 1991)
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A Laboratory Guide to In Vitro Studies of Protein-DNA Interactions (Paperback, Softcover reprint of the original 1st ed. 1991)
Series: Biomethods
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A Safety Considerations Many techniques described here involve a
number of hazards, such as high electrical current and voltage,
radioactivity and highly toxic chemicals. It is absolutely
essential that the instructions of equipment manufacturers be
followed, and that particular attention be paid to the local and
federal safety regulations. B Introduction The expression of
prokaryotic and eukaryotic genes has been shown most often to be
regulated at the level of mRNA synthesis. Thanks to the rapid
development of methods for dissecting DNA sequences, cis-acting
regulatory elements such as promoters and enhancers have been
recognised. More recently, the widely expressed intuition that
discrete sequences within these elements constitute binding sites
for sequence-specific binding proteins has been confirmed,
especially through the use of "footprinting" assays (for examples,
Galas and Schmitz, 1978). This and similar assays have already
resulted in the recognition, isolation and analysis of DNA-bind ing
proteins for several genes. Excellent reviews exist of the
structural studies on these transcription regulatory proteins and
related DNA elements (for example, Glover, 1989 and Johnson and
McKnight, 1989), to which the reader is referred for detailed
information. To set the scene for applications of the techniques
described in this volume, only the barest outline of previous
studies is presented here. Protein-DNA interactions are dependent
on very specific tertiary configurations of the binding protein
which allow the closest contact with the DNA helix.
General
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