Imperial College London

ProfessorPaulFreemont

Faculty of MedicineDepartment of Medicine

Chair in Protein Crystallography
 
 
 
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Contact

 

+44 (0)20 7594 5327p.freemont

 
 
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Location

 

259Sir 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}
}

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 -