Design and engineering genetic tools for Desulfovibrio alaskensis
dc.contributor.advisor
Horsfall, Louise
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dc.contributor.advisor
French, Chris
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dc.contributor.author
Cueva, Miguel Eugenio
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dc.contributor.sponsor
Biotechnology and Biological Sciences Research Council (BBSRC)
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dc.date.accessioned
2020-03-16T11:34:51Z
dc.date.available
2020-03-16T11:34:51Z
dc.date.issued
2020-07-04
dc.description.abstract
Microorganisms, such as the anaerobic bacterium Desulfovibrio alaskensis, have
evolved various mechanisms to resist high concentrations of toxic heavy metals; one
of these mechanisms involves the synthesis of nanoparticles (NPs). It may be
possible to utilise this ability to both reclaim heavy metals from contaminated
effluents and to convert them into industrially useful NPs. By engineering a
genetically modified D. alaskensis, through synthetic biology, cell surface
engineering and by designing a modular cloning (MoClo) toolkit, there is a further
opportunity to tailor nanoparticle synthesis.
DNA assembly techniques have revolutionised biotechnology research and
innovation. However, despite many advances in molecular biology, the assembly of
DNA parts into new constructs remains cumbersome and unpredictable. The
innovation of cloning toolkits and standards such as MoClo have standardised the
process of DNA assembly, making it easier, faster, modular and cost-effective. The
D. alaskensis MoClo toolkit developed in this work consists of characterised oxygen-independent
reporters, synthetic promoters and ribosome binding site (RBS)
libraries.
The D. alaskensis MoClo toolkit was utilised to assemble a combinatorial library of
transcriptional units (TUs) expressing the NiFe hydrogenase small subunit. Platinum
NPs were synthesised by the combinatorial library, and examined for their oxidative
and reduction catalytic activities were tested.
To enhance D. alaskensis resistance to Cu, Pt and Pd, cell surface engineering was
used to express synthetic phytochelatin EC20 on the outer membrane. Tests of
Escherichia coli expressing EC20/IgA to Cu, Pt and Pd concluded that EC20 confers
a higher resistance to all metals.
en
dc.identifier.uri
https://hdl.handle.net/1842/36881
dc.identifier.uri
http://dx.doi.org/10.7488/era/182
dc.language.iso
en
dc.publisher
The University of Edinburgh
en
dc.relation.hasversion
Cueva, M. E., and Horsfall, L. E. (2017) The contribution of microbially produced nanoparticles to sustainable development goals, Microb Biotechnol 10, 1212-1215.
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dc.subject
Desulfovibrio alaskensis
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dc.subject
nanoparticles
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dc.subject
modular cloning
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dc.subject
MoClo toolkit
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dc.subject
oxygen-independent reporters
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dc.subject
synthetic promoters
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dc.subject
ribosome binding site libraries
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dc.title
Design and engineering genetic tools for Desulfovibrio alaskensis
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dc.type
Thesis or Dissertation
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dc.type.qualificationlevel
Doctoral
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dc.type.qualificationname
PhD Doctor of Philosophy
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