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@article{Davenport:2022:10.1049/enb2.12021,
author = {Davenport, B and Tica, J and Isalan, M},
doi = {10.1049/enb2.12021},
journal = {Engineering Biology},
pages = {50--61},
title = {Reducing metabolic burden in the PACEmid evolver system by remastering high copy phagemid vectors},
url = {http://dx.doi.org/10.1049/enb2.12021},
volume = {6},
year = {2022}
}

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TY  - JOUR
AB  - Orthogonal or non-cross-reacting transcription factors are used in synthetic biology as componentsof genetic circuits. Brödel et al. (2016) engineered 12 such cIλ transcription factor variants, using adirected evolution ‘PACEmid’ system. The variants operate as dual activator/repressors and expandgene circuit construction possibilities. However, the high-copy phagemid vectors carrying the cIλvariants imposed high metabolic burden upon cells. Here, we ‘remaster’ the phagemid backbones torelieve their burden substantially, exhibited by a recovery in E. coli growth. The remasteredphagemids’ ability to function within the PACEmid evolver system is maintained, as is the cIλtranscription factors’ activity within these vectors. The low-burden phagemid versions are moresuitable for use in PACEmid experiments and synthetic gene circuits; we have therefore replaced theoriginal high-burden phagemids on the Addgene repository. Our work emphasises the importance ofunderstanding metabolic burden and incorporating it into design steps in future synthetic biologyventures.
AU  - Davenport,B
AU  - Tica,J
AU  - Isalan,M
DO  - 10.1049/enb2.12021
EP  - 61
PY  - 2022///
SN  - 2398-6182
SP  - 50
TI  - Reducing metabolic burden in the PACEmid evolver system by remastering high copy phagemid vectors
T2  - Engineering Biology
UR  - http://dx.doi.org/10.1049/enb2.12021
UR  - http://hdl.handle.net/10044/1/96939
VL  - 6
ER  -

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