Functionality of the β/six Site-Specific Recombination System in Tobacco and Arabidopsis: A Novel Tool for Genetic Engineering of Plant Genomes

Functionality of the β/six Site-Specific Recombination System in Tobacco and Arabidopsis: A... The β recombinase is a member of the prokaryotic site-specific serine recombinases (invertase/resolvase family), which in the presence of a DNA bending cofactor can catalyse DNA deletions between two directly oriented 90-bp six recombination sites. We have examined here whether the β recombinase can be expressed in plants and whether it displays in planta its specific catalytic activity excising DNA sequences that are flanked by six sites. In plant protoplasts, the enzyme could be expressed as a GFP-β recombinase fusion which can localise to the cell nucleus. β recombinase stably expressed in tobacco plants can catalyse deletion of a spacer region that is flanked by directly oriented six sites and has been placed between promoter and a GUS reporter gene (preventing GUS expression). In transient transformation experiments, β recombinase-mediated elimination of the spacer results in transcriptional induction of the GUS gene. Similarly, β recombinase in stably double-transformed Arabidopsis plants deletes specifically the spacer region of a reporter construct that has been incorporated into the genome. In the segregating T1 generation, plants were identified that contain exclusively the recombined reporter construct. In summary, our results demonstrate that functional β recombinase can be expressed in plants and that the enzyme is suitable to precisely eliminate undesired sequences from plant genomes. Therefore, the β/six recombination system (and presumably related recombinases) may become an attractive tool for plant genetic engineering. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Functionality of the β/six Site-Specific Recombination System in Tobacco and Arabidopsis: A Novel Tool for Genetic Engineering of Plant Genomes

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Publisher
Springer Journals
Copyright
Copyright © 2006 by Springer Science+Business Media B.V.
Subject
Life Sciences; Plant Pathology; Biochemistry, general; Plant Sciences
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1007/s11103-006-9108-9
Publisher site
See Article on Publisher Site

Abstract

The β recombinase is a member of the prokaryotic site-specific serine recombinases (invertase/resolvase family), which in the presence of a DNA bending cofactor can catalyse DNA deletions between two directly oriented 90-bp six recombination sites. We have examined here whether the β recombinase can be expressed in plants and whether it displays in planta its specific catalytic activity excising DNA sequences that are flanked by six sites. In plant protoplasts, the enzyme could be expressed as a GFP-β recombinase fusion which can localise to the cell nucleus. β recombinase stably expressed in tobacco plants can catalyse deletion of a spacer region that is flanked by directly oriented six sites and has been placed between promoter and a GUS reporter gene (preventing GUS expression). In transient transformation experiments, β recombinase-mediated elimination of the spacer results in transcriptional induction of the GUS gene. Similarly, β recombinase in stably double-transformed Arabidopsis plants deletes specifically the spacer region of a reporter construct that has been incorporated into the genome. In the segregating T1 generation, plants were identified that contain exclusively the recombined reporter construct. In summary, our results demonstrate that functional β recombinase can be expressed in plants and that the enzyme is suitable to precisely eliminate undesired sequences from plant genomes. Therefore, the β/six recombination system (and presumably related recombinases) may become an attractive tool for plant genetic engineering.

Journal

Plant Molecular BiologySpringer Journals

Published: Nov 28, 2006

References

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