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Transgenic Res ; 21(3): 645-54, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21947784

RESUMO

Trait genes are usually introduced into the plant genome together with a marker gene. The last one becomes unnecessary after transgene selection and characterization. One of the strategies to produce transgenic plants free from the selectable marker is based on site-specific recombination. The present study employed the transient Cre-lox system to remove the nptII marker gene from potato. Transient marker gene excision involves introduction of Cre protein in lox-target plants by PVX virus vector followed by plant regeneration. Using optimized experimental conditions, such as particle bombardment infection method and application of P19 silencing suppressor protein, 20-27% of regenerated plants were identified by PCR analysis as marker-free. Based on our comparison of the recombination frequencies observed in this study to the efficiency of other methods to avoid or eliminate marker genes in potato, we suggest that PVX-Cre mediated site-specific excisional recombination is a useful tool to generate potato plants without superfluous transgenic sequences.


Assuntos
Vetores Genéticos/genética , Integrases/metabolismo , Potexvirus/genética , Solanum tuberosum/metabolismo , Agrobacterium tumefaciens/genética , Southern Blotting , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismo , DNA de Plantas/genética , Inativação Gênica , Genes Reporter , Genes Supressores , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Integrases/genética , Folhas de Planta/genética , Folhas de Planta/metabolismo , Folhas de Planta/virologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plasmídeos/genética , Recombinação Genética , Solanum tuberosum/genética , Tombusvirus/genética , Proteínas Virais/genética , Proteínas Virais/metabolismo
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