Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA sequences, including whole metabolic pathways, onto the Escherichia coli chromosome at well-characterised loci

Suriana Sabri; Jennifer A. Steen; Mareike Bongers; Lars K. Nielsen; Claudia E. Vickers

doi:10.1186/1475-2859-12-60

Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA sequences, including whole metabolic pathways, onto the Escherichia coli chromosome at well-characterised loci

Suriana Sabri, Jennifer A. Steen, Mareike Bongers, Lars K. Nielsen, Claudia E. Vickers

University of Queensland

Research output: Contribution to journal › Journal article › Research › peer-review

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Abstract

Metabolic engineering projects often require integration of multiple genes in order to control the desired phenotype. However, this often requires iterative rounds of engineering because many current insertion approaches are limited by the size of the DNA that can be transferred onto the chromosome. Consequently, construction of highly engineered strains is very time-consuming. A lack of well-characterised insertion loci is also problematic.

Original language	English
Article number	60
Journal	Microbial Cell Factories
Volume	12
Issue number	1
Number of pages	15
ISSN	1475-2859
DOIs	https://doi.org/10.1186/1475-2859-12-60
Publication status	Published - 2013
Externally published	Yes

Bibliographical note

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Keywords

Chromosomal integration
Homologous recombination
Plasmid
Recombineering
E. coli
Xylanase
GFP
Csc genes

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10.1186/1475-2859-12-60

KIKOFinal published version, 819 KB

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title = "Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA sequences, including whole metabolic pathways, onto the Escherichia coli chromosome at well-characterised loci",

abstract = "Metabolic engineering projects often require integration of multiple genes in order to control the desired phenotype. However, this often requires iterative rounds of engineering because many current insertion approaches are limited by the size of the DNA that can be transferred onto the chromosome. Consequently, construction of highly engineered strains is very time-consuming. A lack of well-characterised insertion loci is also problematic.",

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note = "This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.",

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doi = "10.1186/1475-2859-12-60",

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Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA sequences, including whole metabolic pathways, onto the Escherichia coli chromosome at well-characterised loci. / Sabri, Suriana; Steen, Jennifer A.; Bongers, Mareike et al.
In: Microbial Cell Factories, Vol. 12, No. 1, 60, 2013.

Research output: Contribution to journal › Journal article › Research › peer-review

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AU - Sabri, Suriana

AU - Steen, Jennifer A.

AU - Bongers, Mareike

AU - Nielsen, Lars K.

AU - Vickers, Claudia E.

N1 - This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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AB - Metabolic engineering projects often require integration of multiple genes in order to control the desired phenotype. However, this often requires iterative rounds of engineering because many current insertion approaches are limited by the size of the DNA that can be transferred onto the chromosome. Consequently, construction of highly engineered strains is very time-consuming. A lack of well-characterised insertion loci is also problematic.

KW - Chromosomal integration

KW - Homologous recombination

KW - Plasmid

KW - Recombineering

KW - E. coli

KW - Xylanase

KW - GFP

KW - Csc genes

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DO - 10.1186/1475-2859-12-60

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ER -

Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA sequences, including whole metabolic pathways, onto the Escherichia coli chromosome at well-characterised loci

Abstract

Bibliographical note

Keywords

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