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

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...

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Bibliographic Details
Published in:Plant molecular biology 2007-03, Vol.63 (4), p.545
Main Authors: Grønlund, Jesper T, Stemmer, Christian, Lichota, Jacek, Merkle, Thomas, Grasser, Klaus D
Format: Article
Language:English
Subjects:
Deoxyribonucleic acid
DNA
Genetic engineering
Genomics
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Summary: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.[PUBLICATION ABSTRACT]
ISSN:0167-4412
1573-5028
DOI:10.1007/s11103-006-9108-9