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Sakhno L.A., Sytnik E.S., Komarnitskiy I.K., Kuchuk N.V., Klimyuk V.I.
Activity of the Maize Spm Transposon System in Transgenic Orychophragmus violaceus (L.) O.E.Schulz. Plants Obtained by Both Direct Transfer of DNA to Protoplasts and Agrobacterial Transformation of Root Explants
Cytology and Genetics 2002, v 36, number 6, 3-8
Transposon mediated insertional mutagenesis is one of the approaches for the unique gene cloning. A wild species of Cruciferae family Orychophragmus violaceus (L.) O.E.Schulz, which is of interest for practical breeding as a donor of improved plant oil, was an object of the investigation. Plasmid construction used in the experiments included selective NPT II gene, reported GUS gene serving as an excision marker, structural BAR gene located within the dSpm element and Spm transposase. The GUS gene of this plasmid had not his own promoter and became functional only after Spm–transposition. Transformed Orychophragmus violaceus (L.) O.E.Schulz. plants were obtained by direct mesophyll protoplast transformation as well as Agrobacterium tumefaciens-mediated root explant transformation. Gene transfer and the transposition event were confirmed by the GUS activity and the PCR analysis. Relative transformation efficiency using protoplasts was 5.8 %.
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