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Transgenic Res ; 21(3): 645-54, 2012 Jun.
Article in English | MEDLINE | ID: mdl-21947784

ABSTRACT

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.


Subject(s)
Genetic Vectors/genetics , Integrases/metabolism , Potexvirus/genetics , Solanum tuberosum/metabolism , Agrobacterium tumefaciens/genetics , Blotting, Southern , Capsid Proteins/genetics , Capsid Proteins/metabolism , DNA, Plant/genetics , Gene Silencing , Genes, Reporter , Genes, Suppressor , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Integrases/genetics , Plant Leaves/genetics , Plant Leaves/metabolism , Plant Leaves/virology , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Plasmids/genetics , Recombination, Genetic , Solanum tuberosum/genetics , Tombusvirus/genetics , Viral Proteins/genetics , Viral Proteins/metabolism
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