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The Geobacillus plasmid set: a modular toolkit for thermophile engineering

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Title: The Geobacillus plasmid set: a modular toolkit for thermophile engineering
Authors: Reeve, B
Martinez Klimova, E
De Jonghe, J
Leak, DJ
Ellis, T
Item Type: Journal Article
Abstract: Geobacillus thermoglucosidasius is a gram-positive thermophile of industrial interest that exhibits rapid growth and can utilize a variety of plant-derived feedstocks. It is an attractive chassis organism for high temperature biotechnology and synthetic biology applications but is currently limited by a lack of available genetic tools. Here we describe a set of modular shuttle vectors, including a promoter library and reporter proteins. The compact plasmids are composed of interchangeable modules for molecular cloning in Escherichia coli and stable propagation in G. thermoglucosidasius and other Geobacillus species. Modules include two origins of replication, two selectable markers and three reporter proteins for characterization of gene expression. For fine-tuning heterologous expression from these plasmids, we include a characterized promoter library and test ribosome binding site design. Together, these gene expression tools and a standardized plasmid set can facilitate modularity and part exchange to make Geobacillus a thermophile chassis for synthetic biology.
Issue Date: 22-Jun-2016
Date of Acceptance: 22-Jun-2016
URI: http://hdl.handle.net/10044/1/33966
DOI: https://dx.doi.org/10.1021/acssynbio.5b00298
ISSN: 2161-5063
Publisher: American Chemical Society
Start Page: 1342
End Page: 1347
Journal / Book Title: ACS Synthetic Biology
Volume: 5
Issue: 12
Copyright Statement: © 2016 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
Sponsor/Funder: Engineering & Physical Science Research Council (EPSRC)
TMO Renewables Limited
Biotechnology and Biological Sciences Research Cou
Biotechnology and Biological Sciences Research Cou
Engineering & Physical Science Research Council (E
Funder's Grant Number: EP/G036004/1
NA
BB/J001120/1
BB/I00534X/1
EP/J02175X/1
Keywords: Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Biochemistry & Molecular Biology
thermophile
plasmid
Geobacillus
genetic tools
GREEN FLUORESCENT PROTEIN
SYNTHETIC BIOLOGY
ESCHERICHIA-COLI
BACILLUS-STEAROTHERMOPHILUS
NUCLEOTIDE-SEQUENCE
GENE-EXPRESSION
COPY NUMBER
TRANSFORMATION
THERMOGLUCOSIDASIUS
TEMPERATURES
Publication Status: Published
Appears in Collections:Faculty of Engineering
Bioengineering



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