Simple generation of site-directed point mutations in the Escherichia coli chromosome using Red®/ET® recombination

Publikationstyp:
Zeitschriftenaufsatz
Metadaten:
Autoren
  • Ralf Heermann
  • Tim Zeppenfeld
  • Kirsten Jung
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000256151500001&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1186/1475-2859-7-14
Externe Identifier
  • Clarivate Analytics Document Solution ID: 305AZ
  • PubMed Identifier: 18435843
ISSN
1475-2859
Zeitschrift
MICROBIAL CELL FACTORIES
Artikelnummer
ARTN 14
Datum der Veröffentlichung
2008
Status
Published
Titel
Simple generation of site-directed point mutations in the <i>Escherichia coli</i> chromosome using Red®/ET® recombination
Sub types
  • Article
Ausgabe der Zeitschrift
7

Datenquelle: Web of Science (Lite)

Andere Metadatenquellen:
Autoren
  • Ralf Heermann
  • Tim Zeppenfeld
  • Kirsten Jung
DOI
10.1186/1475-2859-7-14
ISSN
1475-2859
Ausgabe der Veröffentlichung
1
Zeitschrift
Microbial Cell Factories
Sprache
en
Paginierung
14 - 14
Datum der Veröffentlichung
2008
Status
Published
Herausgeber
Springer Science and Business Media LLC
Herausgeber URL
http://dx.doi.org/10.1186/1475-2859-7-14
Datum der Datenerfassung
2016
Titel
Simple generation of site-directed point mutations in the Escherichia coli chromosome using Red®/ET® Recombination
Ausgabe der Zeitschrift
7

Datenquelle: Crossref

Abstract
<h4>Background</h4>Introducing point mutations into bacterial chromosomes is important for further progress in studies relying on functional genomics, systems- and synthetic biology, and for metabolic engineering. For many investigations, chromosomal systems are required rather than artificial plasmid based systems.<h4>Results</h4>Here we describe the introduction of a single point mutation into the Escherichia coli chromosome by site-directed mutagenesis without leaving any selection marker. We used Red(R)/ET(R) Recombination in combination with rpsL counter-selection to introduce a single point mutation into the E. coli MG1655 genome, one of the widely used bacterial model strains in systems biology. The method we present is rapid and highly efficient. Since single-stranded synthetic oligonucleotides can be used for recombination, any chromosomal modification can be designed.<h4>Conclusion</h4>Chromosomal modifications performed by rpsL counter-selection may also be used for other bacteria that contain an rpsL homologue, since Red(R)/ET(R) Recombination has been applied to several enteric bacteria before.
Addresses
  • Ludwig-Maximilians-Universität München, Department Biologie I, Bereich Mikrobiologie, Maria-Ward-Str, 1a, D-80638 München, Germany. heermann@lmu.de.
Autoren
  • Ralf Heermann
  • Tim Zeppenfeld
  • Kirsten Jung
DOI
10.1186/1475-2859-7-14
eISSN
1475-2859
Externe Identifier
  • PubMed Identifier: 18435843
  • PubMed Central ID: PMC2373285
Open access
true
ISSN
1475-2859
Zeitschrift
Microbial cell factories
Sprache
eng
Medium
Electronic
Online publication date
2008
Open access status
Open Access
Paginierung
14
Datum der Veröffentlichung
2008
Status
Published
Publisher licence
CC BY
Datum der Datenerfassung
2008
Titel
Simple generation of site-directed point mutations in the Escherichia coli chromosome using Red(R)/ET(R) Recombination.
Sub types
  • research-article
  • Journal Article
Ausgabe der Zeitschrift
7

Files

https://microbialcellfactories.biomedcentral.com/track/pdf/10.1186/1475-2859-7-14 https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/18435843/pdf/?tool=EBI http://www.biomedcentral.com/content/pdf/1475-2859-7-14.pdf https://europepmc.org/articles/PMC2373285?pdf=render

Datenquelle: Europe PubMed Central

Abstract
BACKGROUND: Introducing point mutations into bacterial chromosomes is important for further progress in studies relying on functional genomics, systems- and synthetic biology, and for metabolic engineering. For many investigations, chromosomal systems are required rather than artificial plasmid based systems. RESULTS: Here we describe the introduction of a single point mutation into the Escherichia coli chromosome by site-directed mutagenesis without leaving any selection marker. We used Red(R)/ET(R) Recombination in combination with rpsL counter-selection to introduce a single point mutation into the E. coli MG1655 genome, one of the widely used bacterial model strains in systems biology. The method we present is rapid and highly efficient. Since single-stranded synthetic oligonucleotides can be used for recombination, any chromosomal modification can be designed. CONCLUSION: Chromosomal modifications performed by rpsL counter-selection may also be used for other bacteria that contain an rpsL homologue, since Red(R)/ET(R) Recombination has been applied to several enteric bacteria before.
Date of acceptance
2008
Autoren
  • Ralf Heermann
  • Tim Zeppenfeld
  • Kirsten Jung
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/18435843
DOI
10.1186/1475-2859-7-14
eISSN
1475-2859
Externe Identifier
  • PubMed Central ID: PMC2373285
Zeitschrift
Microb Cell Fact
Sprache
eng
Country
England
Paginierung
14
PII
1475-2859-7-14
Datum der Veröffentlichung
2008
Status
Published online
Datum, an dem der Datensatz öffentlich gemacht wurde
2012
Titel
Simple generation of site-directed point mutations in the Escherichia coli chromosome using Red(R)/ET(R) Recombination.
Sub types
  • Journal Article
Ausgabe der Zeitschrift
7

Datenquelle: PubMed

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