PCR assembly of synthetic promoters

Hodan Mohamed, David Gould*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

In this chapter, we describe a two-step assembly PCR method to construct synthetic promoters. Essentially, this method takes advantage of specific annealing between complimentary DNA sequences to build random TFBS combinations within the assembled PCR products. A DNA polymerase is then employed to fill in the unpaired nucleotides in the generated sequences and also to amplify the assembled PCR products. We have used this method to generate synthetic promoters whereby the orientation of the TFBS can be controlled, the spacing between TFBS can be predetermined, and also the full diversity of the consensus TFBS can be covered through the use of degenerate oligonucleotides.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages147-156
Number of pages10
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1651
ISSN (Print)1064-3745

Bibliographical note

Publisher Copyright:
© 2017, Springer Science+Business Media LLC.

Keywords

  • Assembly PCR
  • Multi-responsive promoters
  • Random assembly
  • Synthetic promoters

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