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  • Open Access

Promoter analysis by saturation mutagenesis

Biological Procedures Online3:31064

  • Received: 30 August 2001
  • Accepted: 17 December 2001


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