Imperial College London
Alternate

DrJohnHeap

Faculty of Natural Sciences, Department of Life Sciences

Honorary Senior Lecturer
 
 
 
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Contact

 

+44 (0)20 7594 5355j.heap

 
 
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Location

 

Bessemer BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Kelly:2016:10.1021/acssynbio.6b00030,
author = {Kelly, CL and Liu, Z and Yoshihara, A and Jenkinson, SF and Wormald, MR and Otero, J and Estévez, A and Kato, A and Marqvorsen, MHS and Fleet, GWJ and Estévez, RJ and Izumori, K and Heap, JT},
doi = {10.1021/acssynbio.6b00030},
journal = {ACS Synth Biol},
pages = {1136--1145},
title = {Synthetic Chemical Inducers and Genetic Decoupling Enable Orthogonal Control of the rhaBAD Promoter.},
url = {http://dx.doi.org/10.1021/acssynbio.6b00030},
volume = {5},
year = {2016}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - External control of gene expression is crucial in synthetic biology and biotechnology research and applications, and is commonly achieved using inducible promoter systems. The E. coli rhamnose-inducible rhaBAD promoter has properties superior to more commonly used inducible expression systems, but is marred by transient expression caused by degradation of the native inducer, l-rhamnose. To address this problem, 35 analogues of l-rhamnose were screened for induction of the rhaBAD promoter, but no strong inducers were identified. In the native configuration, an inducer must bind and activate two transcriptional activators, RhaR and RhaS. Therefore, the expression system was reconfigured to decouple the rhaBAD promoter from the native rhaSR regulatory cascade so that candidate inducers need only activate the terminal transcription factor RhaS. Rescreening the 35 compounds using the modified rhaBAD expression system revealed several promising inducers. These were characterized further to determine the strength, kinetics, and concentration-dependence of induction; whether the inducer was used as a carbon source by E. coli; and the modality (distribution) of induction among populations of cells. l-Mannose was found to be the most useful orthogonal inducer, providing an even greater range of induction than the native inducer l-rhamnose, and crucially, allowing sustained induction instead of transient induction. These findings address the key limitation of the rhaBAD expression system and suggest it may now be the most suitable system for many applications.
AU  - Kelly,CL
AU  - Liu,Z
AU  - Yoshihara,A
AU  - Jenkinson,SF
AU  - Wormald,MR
AU  - Otero,J
AU  - Estévez,A
AU  - Kato,A
AU  - Marqvorsen,MHS
AU  - Fleet,GWJ
AU  - Estévez,RJ
AU  - Izumori,K
AU  - Heap,JT
DO  - 10.1021/acssynbio.6b00030
EP  - 1145
PY  - 2016///
SP  - 1136
TI  - Synthetic Chemical Inducers and Genetic Decoupling Enable Orthogonal Control of the rhaBAD Promoter.
T2  - ACS Synth Biol
UR  - http://dx.doi.org/10.1021/acssynbio.6b00030
UR  - https://www.ncbi.nlm.nih.gov/pubmed/27247275
UR  - http://hdl.handle.net/10044/1/33313
VL  - 5
ER  -
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