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PMID: 17644634 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Directed evolution of AraC for improved compatibility of arabinose- and lactose-inducible promoters.

Applied and environmental microbiology ·Vol. 73 ·No. 18 ·2007-09-00 ·Pages 5711-5

Lee SK, Chou HH, Pfleger BF, Newman JD, Yoshikuni Y, Keasling JD

Abstract

Synthetic biological systems often require multiple, independently inducible promoters in order to control the expression levels of several genes; however, cross talk between the promoters limits this ability. Here, we demonstrate the directed evolution of AraC to construct an arabinose-inducible (P(BAD)) system that is more compatible with IPTG (isopropyl-beta-D-1-thiogalactopyranoside) induction of a lactose-inducible (P(lac)) system. The constructed system is 10 times more sensitive to arabinose and tolerates IPTG significantly better than the wild type. Detailed studies indicate that the AraC dimerization domain and C terminus are important for the increased sensitivity of AraC to arabinose.

MeSH Terms
AraC Transcription Factor/drug effects,genetics Arabinose/metabolism,pharmacology Escherichia coli/genetics,growth & development,metabolism Escherichia coli Proteins/drug effects,genetics Evolution, Molecular Isopropyl Thiogalactoside/metabolism,pharmacology Lactose/pharmacology Mutagenesis, Site-Directed Operon Promoter Regions, Genetic/drug effects,physiology
Chemicals
AraC Transcription Factor AraC protein, E coli Escherichia coli Proteins Isopropyl Thiogalactoside Arabinose Lactose
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lee Sung Kuk
Department of Chemical Engineering, University of California, Berkeley, California 94720, USA.
Chou Howard H
Pfleger Brian F
Newman Jack D
Yoshikuni Yasuo
Keasling Jay D
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.00791-07
Published
2007-09-00
Epub
2007-00-20
Pages
5711-5
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC2074931
Subset
IM
Grants
NIGMS NIH HHS · R01 GM070763 · United States
NIGMS NIH HHS · GM070763-01 · United States
External Links
PubMed source DOI: 10.1128/AEM.00791-07 PMC Full Text View journal details
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