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ParA resolvase catalyzes site-specific excision of DNA from the Arabidopsis genome
Authors:James G. Thomson  Yuan-Yeu Yau  Robert Blanvillain  Dawn Chiniquy  Roger Thilmony  David W. Ow
Affiliation:(1) Plant Gene Expression Center, 800 Buchanan Street, Albany, CA 94710, USA;(2) Department of Plant and Microbial Biology, University of California at Berkeley, Berkeley, CA 94720, USA;(3) Crop Improvement and Utilization Research Unit, Western Regional Research Center, USDA-ARS, 800 Buchanan Street, Albany, CA 94710, USA;(4) Department of Plant Biology, University of California at Davis, Davis, CA 95616, USA
Abstract:The small serine resolvase ParA from bacterial plasmids RK2 and RP4 catalyzes the recombination of two identical 133 bp recombination sites known as MRS. Previously, we reported that ParA is active in the fission yeast Schizosaccharomyces pombe. In this work, the parA recombinase gene was placed under the control of the Arabidopsis OXS3 promoter and introduced into Arabidopsis lines harboring a chromosomally integrated MRS-flanked target. The ParA recombinase excised the MRS-flanked DNA and the excision event was detected in subsequent generations in the absence of ParA, indicating germinal transmission of the excision event. The precise site-specific deletion by the ParA recombination system in planta demonstrates that the ParA recombinase can be used to remove transgenic DNA, such as selectable markers or other introduced transgenes that are no longer desired in the final product.
Keywords:Cre-lox   Marker excision  Genetic engineering   OXS3   Site-specific recombination
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