title: Utilization of PVX-Cre expression vector in potato.
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.
authors.affiliation: Julius Kuehn Institute, Federal Research Centre for Cultivated Plants (JKI), Institute for Biosafety of Genetically Modified Plants, Erwin-Baur-Str 27, 06484 Quedlinburg, Germany.
authors.forename: Lilya
authors.initials: L
authors.lastname: Kopertekh
authors.forename: Veronica v
authors.initials: Vv
authors.lastname: Saint Paul
authors.forename: Erika
authors.initials: E
authors.lastname: Krebs
authors.forename: Joachim
authors.initials: J
authors.lastname: Schiemann
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D015139:Blotting, Southern; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D018744:DNA, Plant / Q000235:genetics; D020868:Gene Silencing; D017930:Genes, Reporter; D016153:Genes, Suppressor; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000821:virology; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D017863:Potexvirus / Q000235:genetics*; D011995:Recombination, Genetic; D011198:Solanum tuberosum / Q000235:genetics / Q000378:metabolism*; D017864:Tombusvirus / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D018744:DNA, Plant; C493885:P19 protein, tomato bushy stunt virus; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases