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Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome
Direct manipulation of bacterial chromosomes by recombination-based techniques has become increasingly important for both cognitive and applied research. Here we demonstrate, for the first time, the combination of the Red recombinase system with I-SceI endonuclease-based selection of successful reco...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3021506/ https://www.ncbi.nlm.nih.gov/pubmed/21264289 http://dx.doi.org/10.1371/journal.pone.0015763 |
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author | Blank, Kathrin Hensel, Michael Gerlach, Roman G. |
author_facet | Blank, Kathrin Hensel, Michael Gerlach, Roman G. |
author_sort | Blank, Kathrin |
collection | PubMed |
description | Direct manipulation of bacterial chromosomes by recombination-based techniques has become increasingly important for both cognitive and applied research. Here we demonstrate, for the first time, the combination of the Red recombinase system with I-SceI endonuclease-based selection of successful recombinants after electroporation with short synthetic olignucleotides. We show the generation of scarless gene knockouts as well as site-directed mutagenesis using the Salmonella virulence-associated two component signaling system PhoPQ. The presented approach is very versatile for generating in-frame deletions, point mutations or insertions within bacterial chromosomes. |
format | Text |
id | pubmed-3021506 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-30215062011-01-24 Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome Blank, Kathrin Hensel, Michael Gerlach, Roman G. PLoS One Research Article Direct manipulation of bacterial chromosomes by recombination-based techniques has become increasingly important for both cognitive and applied research. Here we demonstrate, for the first time, the combination of the Red recombinase system with I-SceI endonuclease-based selection of successful recombinants after electroporation with short synthetic olignucleotides. We show the generation of scarless gene knockouts as well as site-directed mutagenesis using the Salmonella virulence-associated two component signaling system PhoPQ. The presented approach is very versatile for generating in-frame deletions, point mutations or insertions within bacterial chromosomes. Public Library of Science 2011-01-14 /pmc/articles/PMC3021506/ /pubmed/21264289 http://dx.doi.org/10.1371/journal.pone.0015763 Text en Blank et al. 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 Blank, Kathrin Hensel, Michael Gerlach, Roman G. Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome |
title | Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome |
title_full | Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome |
title_fullStr | Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome |
title_full_unstemmed | Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome |
title_short | Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome |
title_sort | rapid and highly efficient method for scarless mutagenesis within the salmonella enterica chromosome |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3021506/ https://www.ncbi.nlm.nih.gov/pubmed/21264289 http://dx.doi.org/10.1371/journal.pone.0015763 |
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