Protocol: Precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli.

Laura C Roden; Berthold  Göttgens; Euphemia Mutasa-Gottgens

doi:10.1186/1746-4811-1-6

Protocol: Precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli.

Laura C Roden, Berthold Göttgens, Euphemia Mutasa-Gottgens

School of Life and Medical Sciences

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

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Abstract

Plant genome sequence data now provide opportunities to conduct molecular genetic studies at the level of the whole gene locus and above. Such studies will be greatly facilitated by adopting and developing further the new generation of genetic engineering tools, based on homologous recombination cloning in Escherichia coli, which are free from the constraints imposed by the availability of suitably positioned restriction sites. Here we describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs coordinately driven by locus-specific regulatory elements.

Original language	English
Number of pages	12
Journal	Plant Methods
Volume	1
Issue number	6
Early online date	29 Sept 2005
DOIs	https://doi.org/10.1186/1746-4811-1-6
Publication status	Published - 2005

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10.1186/1746-4811-1-6

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© 2005 Roden et al; licensee 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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abstract = "Plant genome sequence data now provide opportunities to conduct molecular genetic studies at the level of the whole gene locus and above. Such studies will be greatly facilitated by adopting and developing further the new generation of genetic engineering tools, based on homologous recombination cloning in Escherichia coli, which are free from the constraints imposed by the availability of suitably positioned restriction sites. Here we describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs coordinately driven by locus-specific regulatory elements.",

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AU - Göttgens, Berthold

AU - Mutasa-Gottgens, Euphemia

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N2 - Plant genome sequence data now provide opportunities to conduct molecular genetic studies at the level of the whole gene locus and above. Such studies will be greatly facilitated by adopting and developing further the new generation of genetic engineering tools, based on homologous recombination cloning in Escherichia coli, which are free from the constraints imposed by the availability of suitably positioned restriction sites. Here we describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs coordinately driven by locus-specific regulatory elements.

AB - Plant genome sequence data now provide opportunities to conduct molecular genetic studies at the level of the whole gene locus and above. Such studies will be greatly facilitated by adopting and developing further the new generation of genetic engineering tools, based on homologous recombination cloning in Escherichia coli, which are free from the constraints imposed by the availability of suitably positioned restriction sites. Here we describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs coordinately driven by locus-specific regulatory elements.

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Protocol: Precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli.

Abstract

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