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  • Title: Plant regeneration methods for rapid generation of a large scale Ds transposant population in rice.
    Author: Xuan YH, Huang J, Yi G, Park DS, Park SK, Eun MY, Yun DW, Lee GS, Kim TH, Han CD.
    Journal: Methods Mol Biol; 2013; 1057():101-16. PubMed ID: 23918423.
    Abstract:
    To mutagenize rice genomes, a two-element system is utilized. This system comprises an immobile Ac element driven by the CaMV 35S promoter, and a gene trap Ds carrying a partial intron with alternative splice acceptors fused to the GUS coding region. Rapid, large-scale generation of a Ds transposant population was achieved using a plant regeneration procedure involving the tissue culture of seed-derived calli carrying Ac and Ds elements. During tissue cultures, Ds mobility accompanies changes in methylation patterns of a terminal region of Ds, where over 70% of plants contained independent Ds insertions. In the transposon population, around 12% of plants expressed GUS at the early seedling stage. A flanking-sequence-tag (FST) database has been established by cloning over 19,968 Ds insertion sites and the Ds map shows relatively uniform distribution across the rice chromosomes.
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