Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Juhas, Mario, Ajioka, James W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
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
_version_ 1782363783692288000
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
work_keys_str_mv AT juhasmario identificationandvalidationofnovelchromosomalintegrationandexpressionlociinescherichiacoliflagellarregion1
AT ajiokajamesw identificationandvalidationofnovelchromosomalintegrationandexpressionlociinescherichiacoliflagellarregion1