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A roadmap for gene system development in Clostridium
Clostridium species are both heroes and villains. Some cause serious human and animal diseases, those present in the gut microbiota generally contribute to health and wellbeing, while others represent useful industrial chassis for the production of chemicals and fuels. To understand, counter or expl...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Academic Press
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5058259/ https://www.ncbi.nlm.nih.gov/pubmed/27234263 http://dx.doi.org/10.1016/j.anaerobe.2016.05.011 |
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author | Minton, Nigel P. Ehsaan, Muhammad Humphreys, Christopher M. Little, Gareth T. Baker, Jonathan Henstra, Anne M. Liew, Fungmin Kelly, Michelle L. Sheng, Lili Schwarz, Katrin Zhang, Ying |
author_facet | Minton, Nigel P. Ehsaan, Muhammad Humphreys, Christopher M. Little, Gareth T. Baker, Jonathan Henstra, Anne M. Liew, Fungmin Kelly, Michelle L. Sheng, Lili Schwarz, Katrin Zhang, Ying |
author_sort | Minton, Nigel P. |
collection | PubMed |
description | Clostridium species are both heroes and villains. Some cause serious human and animal diseases, those present in the gut microbiota generally contribute to health and wellbeing, while others represent useful industrial chassis for the production of chemicals and fuels. To understand, counter or exploit, there is a fundamental requirement for effective systems that may be used for directed or random genome modifications. We have formulated a simple roadmap whereby the necessary gene systems maybe developed and deployed. At its heart is the use of ‘pseudo-suicide’ vectors and the creation of a pyrE mutant (a uracil auxotroph), initially aided by ClosTron technology, but ultimately made using a special form of allelic exchange termed ACE (Allele-Coupled Exchange). All mutants, regardless of the mutagen employed, are made in this host. This is because through the use of ACE vectors, mutants can be rapidly complemented concomitant with correction of the pyrE allele and restoration of uracil prototrophy. This avoids the phenotypic effects frequently observed with high copy number plasmids and dispenses with the need to add antibiotic to ensure plasmid retention. Once available, the pyrE host may be used to stably insert all manner of application specific modules. Examples include, a sigma factor to allow deployment of a mariner transposon, hydrolases involved in biomass deconstruction and therapeutic genes in cancer delivery vehicles. To date, provided DNA transfer is obtained, we have not encountered any clostridial species where this technology cannot be applied. These include, Clostridium difficile, Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium botulinum, Clostridium perfringens, Clostridium sporogenes, Clostridium pasteurianum, Clostridium ljungdahlii, Clostridium autoethanogenum and even Geobacillus thermoglucosidasius. |
format | Online Article Text |
id | pubmed-5058259 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Academic Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-50582592016-10-17 A roadmap for gene system development in Clostridium Minton, Nigel P. Ehsaan, Muhammad Humphreys, Christopher M. Little, Gareth T. Baker, Jonathan Henstra, Anne M. Liew, Fungmin Kelly, Michelle L. Sheng, Lili Schwarz, Katrin Zhang, Ying Anaerobe Article Clostridium species are both heroes and villains. Some cause serious human and animal diseases, those present in the gut microbiota generally contribute to health and wellbeing, while others represent useful industrial chassis for the production of chemicals and fuels. To understand, counter or exploit, there is a fundamental requirement for effective systems that may be used for directed or random genome modifications. We have formulated a simple roadmap whereby the necessary gene systems maybe developed and deployed. At its heart is the use of ‘pseudo-suicide’ vectors and the creation of a pyrE mutant (a uracil auxotroph), initially aided by ClosTron technology, but ultimately made using a special form of allelic exchange termed ACE (Allele-Coupled Exchange). All mutants, regardless of the mutagen employed, are made in this host. This is because through the use of ACE vectors, mutants can be rapidly complemented concomitant with correction of the pyrE allele and restoration of uracil prototrophy. This avoids the phenotypic effects frequently observed with high copy number plasmids and dispenses with the need to add antibiotic to ensure plasmid retention. Once available, the pyrE host may be used to stably insert all manner of application specific modules. Examples include, a sigma factor to allow deployment of a mariner transposon, hydrolases involved in biomass deconstruction and therapeutic genes in cancer delivery vehicles. To date, provided DNA transfer is obtained, we have not encountered any clostridial species where this technology cannot be applied. These include, Clostridium difficile, Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium botulinum, Clostridium perfringens, Clostridium sporogenes, Clostridium pasteurianum, Clostridium ljungdahlii, Clostridium autoethanogenum and even Geobacillus thermoglucosidasius. Academic Press 2016-10 /pmc/articles/PMC5058259/ /pubmed/27234263 http://dx.doi.org/10.1016/j.anaerobe.2016.05.011 Text en © 2016 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Minton, Nigel P. Ehsaan, Muhammad Humphreys, Christopher M. Little, Gareth T. Baker, Jonathan Henstra, Anne M. Liew, Fungmin Kelly, Michelle L. Sheng, Lili Schwarz, Katrin Zhang, Ying A roadmap for gene system development in Clostridium |
title | A roadmap for gene system development in Clostridium |
title_full | A roadmap for gene system development in Clostridium |
title_fullStr | A roadmap for gene system development in Clostridium |
title_full_unstemmed | A roadmap for gene system development in Clostridium |
title_short | A roadmap for gene system development in Clostridium |
title_sort | roadmap for gene system development in clostridium |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5058259/ https://www.ncbi.nlm.nih.gov/pubmed/27234263 http://dx.doi.org/10.1016/j.anaerobe.2016.05.011 |
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