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Identification and Validation of Novel Chromosomal Integration and Expression Loci in Escherichia coli Flagellar Region 1
Escherichia coli is used as a chassis for a number of Synthetic Biology applications. The lack of suitable chromosomal integration and expression loci is among the main hurdles of the E. coli engineering efforts. We identified and validated chromosomal integration and expression target sites within...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4376774/ https://www.ncbi.nlm.nih.gov/pubmed/25816013 http://dx.doi.org/10.1371/journal.pone.0123007 |
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author | Juhas, Mario Ajioka, James W. |
author_facet | Juhas, Mario Ajioka, James W. |
author_sort | Juhas, Mario |
collection | PubMed |
description | Escherichia coli is used as a chassis for a number of Synthetic Biology applications. The lack of suitable chromosomal integration and expression loci is among the main hurdles of the E. coli engineering efforts. We identified and validated chromosomal integration and expression target sites within E. coli K12 MG1655 flagellar region 1. We analyzed five open reading frames of the flagellar region 1, flgA, flgF, flgG, flgI, and flgJ, that are well-conserved among commonly-used E. coli strains, such as MG1655, W3110, DH10B and BL21-DE3. The efficiency of the integration into the E. coli chromosome and the expression of the introduced genetic circuit at the investigated loci varied significantly. The integrations did not have a negative impact on growth; however, they completely abolished motility. From the investigated E. coli K12 MG1655 flagellar region 1, flgA and flgG are the most suitable chromosomal integration and expression loci. |
format | Online Article Text |
id | pubmed-4376774 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-43767742015-04-04 Identification and Validation of Novel Chromosomal Integration and Expression Loci in Escherichia coli Flagellar Region 1 Juhas, Mario Ajioka, James W. PLoS One Research Article Escherichia coli is used as a chassis for a number of Synthetic Biology applications. The lack of suitable chromosomal integration and expression loci is among the main hurdles of the E. coli engineering efforts. We identified and validated chromosomal integration and expression target sites within E. coli K12 MG1655 flagellar region 1. We analyzed five open reading frames of the flagellar region 1, flgA, flgF, flgG, flgI, and flgJ, that are well-conserved among commonly-used E. coli strains, such as MG1655, W3110, DH10B and BL21-DE3. The efficiency of the integration into the E. coli chromosome and the expression of the introduced genetic circuit at the investigated loci varied significantly. The integrations did not have a negative impact on growth; however, they completely abolished motility. From the investigated E. coli K12 MG1655 flagellar region 1, flgA and flgG are the most suitable chromosomal integration and expression loci. Public Library of Science 2015-03-27 /pmc/articles/PMC4376774/ /pubmed/25816013 http://dx.doi.org/10.1371/journal.pone.0123007 Text en © 2015 Juhas, Ajioka http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Juhas, Mario Ajioka, James W. Identification and Validation of Novel Chromosomal Integration and Expression Loci in Escherichia coli Flagellar Region 1 |
title | Identification and Validation of Novel Chromosomal Integration and Expression Loci in Escherichia coli Flagellar Region 1 |
title_full | Identification and Validation of Novel Chromosomal Integration and Expression Loci in Escherichia coli Flagellar Region 1 |
title_fullStr | Identification and Validation of Novel Chromosomal Integration and Expression Loci in Escherichia coli Flagellar Region 1 |
title_full_unstemmed | Identification and Validation of Novel Chromosomal Integration and Expression Loci in Escherichia coli Flagellar Region 1 |
title_short | Identification and Validation of Novel Chromosomal Integration and Expression Loci in Escherichia coli Flagellar Region 1 |
title_sort | identification and validation of novel chromosomal integration and expression loci in escherichia coli flagellar region 1 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4376774/ https://www.ncbi.nlm.nih.gov/pubmed/25816013 http://dx.doi.org/10.1371/journal.pone.0123007 |
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