Open Access

Promoter analysis by saturation mutagenesis

Biological Procedures Online3:31064

https://doi.org/10.1251/bpo24

Received: 30 August 2001

Accepted: 17 December 2001

Abstract

Gene expression and regulation are mediated by DNA sequences, in most instances, directly upstream to the coding sequences by recruiting transcription factors, regulators, and a RNA polymerase in a spatially defined fashion. Few nucleotides within a promoter make contact with the bound proteins. The minimal set of nucleotides that can recruit a protein factor is called a cis-acting element. This article addresses a powerful mutagenesis strategy that can be employed to define cis-acting elements at a molecular level. Technical details including primer design, saturation mutagenesis, construction of promoter libraries, phenotypic analysis, data analysis, and interpretation are discussed.

Indexing terms

promoter regions (genetics) mutagenesis polymerase chain reaction

Notes

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