Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academic Press 2016
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
_version_ 1782459215308128256
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
work_keys_str_mv AT mintonnigelp aroadmapforgenesystemdevelopmentinclostridium
AT ehsaanmuhammad aroadmapforgenesystemdevelopmentinclostridium
AT humphreyschristopherm aroadmapforgenesystemdevelopmentinclostridium
AT littlegaretht aroadmapforgenesystemdevelopmentinclostridium
AT bakerjonathan aroadmapforgenesystemdevelopmentinclostridium
AT henstraannem aroadmapforgenesystemdevelopmentinclostridium
AT liewfungmin aroadmapforgenesystemdevelopmentinclostridium
AT kellymichellel aroadmapforgenesystemdevelopmentinclostridium
AT shenglili aroadmapforgenesystemdevelopmentinclostridium
AT schwarzkatrin aroadmapforgenesystemdevelopmentinclostridium
AT zhangying aroadmapforgenesystemdevelopmentinclostridium
AT mintonnigelp roadmapforgenesystemdevelopmentinclostridium
AT ehsaanmuhammad roadmapforgenesystemdevelopmentinclostridium
AT humphreyschristopherm roadmapforgenesystemdevelopmentinclostridium
AT littlegaretht roadmapforgenesystemdevelopmentinclostridium
AT bakerjonathan roadmapforgenesystemdevelopmentinclostridium
AT henstraannem roadmapforgenesystemdevelopmentinclostridium
AT liewfungmin roadmapforgenesystemdevelopmentinclostridium
AT kellymichellel roadmapforgenesystemdevelopmentinclostridium
AT shenglili roadmapforgenesystemdevelopmentinclostridium
AT schwarzkatrin roadmapforgenesystemdevelopmentinclostridium
AT zhangying roadmapforgenesystemdevelopmentinclostridium