Imperial College London
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DrRichardKelwick

Faculty of Medicine, Department of Infectious Disease

Research Associate
 
 
 
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Contact

 

+44 (0)20 7594 3058r.kelwick Website

 
 
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Location

 

Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Kelwick:2016:10.1016/j.ymben.2016.09.008,
author = {Kelwick, RJR and Webb, AJ and MacDonald, JT and Freemont, PS},
doi = {10.1016/j.ymben.2016.09.008},
journal = {Metabolic Engineering},
pages = {370--381},
title = {Development of a Bacillus subtilis cell-free transcription-translation system for prototyping regulatory elements},
url = {http://dx.doi.org/10.1016/j.ymben.2016.09.008},
volume = {38},
year = {2016}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Cell-free transcription-translation systems were originally applied towards in vitro protein production. More recently, synthetic biology is enabling these systems to be used within a systematic design context for prototyping DNA regulatory elements, genetic logic circuits and biosynthetic pathways. The Gram-positive soil bacterium, Bacillus subtilis, is an established model organism of industrial importance. To this end, we developed several B. subtilis-based cell-free systems. Our improved B. subtilis WB800N-based system was capable of producing 0.8 µM GFP, which gave a ~72x fold-improvement when compared with a B. subtilis 168 cell-free system. Our improved system was applied towards the prototyping of a B. subtilis promoter library in which we engineered several promoters, derived from the wild-type Pgrac (σA) promoter, that display a range of comparable in vitro and in vivo transcriptional activities. Additionally, we demonstrate the cell-free characterisation of an inducible expression system, and the activity of a model enzyme - renilla luciferase.
AU  - Kelwick,RJR
AU  - Webb,AJ
AU  - MacDonald,JT
AU  - Freemont,PS
DO  - 10.1016/j.ymben.2016.09.008
EP  - 381
PY  - 2016///
SN  - 1096-7184
SP  - 370
TI  - Development of a Bacillus subtilis cell-free transcription-translation system for prototyping regulatory elements
T2  - Metabolic Engineering
UR  - http://dx.doi.org/10.1016/j.ymben.2016.09.008
UR  - http://hdl.handle.net/10044/1/41096
VL  - 38
ER  -
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