Abstract
A number of mutagenesis methods allow the systematic survey of a region of transcriptional regulatory sequence for identifying functional elements. In these methods, clusters or blocks of point mutations are introduced at discrete locations that span a suspected regulatory region of a gene. The individual mutants are then examined for retention of transcriptional activity typically in a reporter gene assay. Traditional methods, including linker-scanning mutagenesis (1) and microscale “shot-gun” gene synthesis (2), are either tedious and time-consuming or relatively inefficient at producing the desired mutated sequence. A modification of the PCR-based mutagenesis method originally described by Li and Shapiro (3) is much simpler, faster, and more efficient than the traditional methods.
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© 1997 Humana Press Inc.
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Schanke, J.T. (1997). Linker Scanning Mutagenesis by Three-Step PCR. In: White, B.A. (eds) PCR Cloning Protocols. Methods in Molecular Biology™, vol 67. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-483-6:197
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DOI: https://doi.org/10.1385/0-89603-483-6:197
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-483-9
Online ISBN: 978-1-59259-553-2
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