PCR assembly of synthetic promoters

Hodan Mohamed; David Gould

doi:10.1007/978-1-4939-7223-4_11

PCR assembly of synthetic promoters

Hodan Mohamed, David Gould^*

^*Corresponding author for this work

Antiviral Susceptibility Reference Unit General

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

1 Citation (Scopus)

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 language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	147-156
Number of pages	10
DOIs	https://doi.org/10.1007/978-1-4939-7223-4_11
Publication status	Published - 2017

Publication series

Name	Methods in Molecular Biology
Volume	1651
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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10.1007/978-1-4939-7223-4_11

Cite this

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KW - Assembly PCR

KW - Multi-responsive promoters

KW - Random assembly

KW - Synthetic promoters

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BT - Methods in Molecular Biology

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ER -

PCR assembly of synthetic promoters

Abstract

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Bibliographical note

Keywords

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