A system to induce the deletion of gemomic sequences using R/RS site-specific recombination and the Ac transposon in transgenic rice plants |
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Authors: | Y Nakagawa C Machida Y Machida K Toriyama |
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Institution: | (1) Laboratory of Plant Breeding and Genetics, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981-8555, Japan e-mail: torikin@bios.tohoku.ac.jp Fax: +81-22-7178654, JP;(2) Laboratory of Developmental Biology, Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan, JP |
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Abstract: | Recombinase encoded by the R gene of Zygosaccharomyces
rouxii mediates reciprocal recombination between two specific recombination sites (RSs) to induce deletion or inversion of the DNA segment that is flanked by the RSs. The R gene under the control of the CaMV 35 S promoter was introduced into rice (Oryza sativa L.). R/RS-specific deletion was first demonstrated in transgenic rice callus carrying the R gene by transient introduction of a cryptic reporter gene that was designed to confer β-glucuronidase (GUS) expression once
deletion between two RSs took place. The rice containing the R gene was subsequently crossed with transgenic rice carrying (I-RS/dAc-I-RS) T-DNA that contained RS sequences within the T-DNA and another RS in a modified Ac element that had been transposed to a new locus by Ac transposase. Deletion of the gemomic sequences flanked by the two RSs was detected by PCR analysis in somatic cells of F2 plants. These results demonstrate a technical advance in that the R/RS recombination system, in combination with the Ac transposable element, can be used to generate deletion in rice chromosomes.
Received: 30 June 2000 / Accepted: 16 October 2000 |
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Keywords: | Deletion Oryza sativa L R/RS recombinationsystem Site-specific recombination Transposon Ac |
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