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Genetic transformation of Medicago species by Agrobacterium tumefaciens and electroporation of protoplasts
Authors:Nicolay Kuchuk  Igor Komarnitski  Anatoliy Shakhovsky  Yuri Gleba
Affiliation:(1) Division of Cell Biology and Engineering, Academy of Sciences of the Ukr. S.S.R., Acad. Lebedeva str. 1, GSP-22, 252650 Kiev, USSR
Abstract:Shoot and leaf segments of a non-regenerable Medicago sativa L. genotype were cocultivated with the ldquoshootyrdquo mutant of Agrobacterium tumefaciens carrying the pGV 2206 plasmid. Transformed callus lines were selected and regenerated on the hormone free B5 medium. Southern blot analysis demonstrated integration of T-DNA in to the genome of the regenerated plants.Transgenic plants resistant to kanamycin were obtained by electroporation of Medicago borealis protoplasts with the pGA 472 plasmid DNA.Abbreviations 2.4 D 2.4 dichlorophenoxyacetic acid - BAP 6-benzyladenine - T-DNA transferred DNA into plants from Ti-plasmid of A. tumefaciens
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