title: High frequency of single-copy T-DNA transformants produced after floral dip in CRE-expressing Arabidopsis plants.
abstract: Transgenic plants that harbor a single copy of the introduced transgene are preferable to those with multiple transgene copies because multiple T-DNA copies correlate with expression variability and susceptibility to silencing. Especially after the commonly used floral-dip Agrobacterium-mediated transformation method, the frequency of single-copy transformants is low. The CRE/loxP recombinase-based strategy to resolve complex T-DNA loci has proven to be successful to efficiently obtain single-copy T-DNA transformants by directly transforming loxP-containing T-DNA vectors in CRE-expressing Arabidopsis thaliana plants. This chapter describes in detail how to transform three available loxP-containing T-DNA vectors into CRE-producing Arabidopsis C24 plants and subsequently how to analyze the transgenic plants for the T-DNA locus structure.
authors.affiliation: Department of Plant Systems Biology, VIB, Department of Biotechnology and Bioinformatics, Ghent University, Ghent, Belgium.
authors.forename: Annelies
authors.initials: A
authors.lastname: De Paepe
authors.forename: Sylvie
authors.initials: S
authors.lastname: De Buck
authors.forename: Jonah
authors.initials: J
authors.lastname: Nolf
authors.forename: Ann
authors.initials: A
authors.lastname: Depicker
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D018628:Gene Dosage; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D019426:Integrases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial; D018744:DNA, Plant; C036483:T-DNA; C045073:Cre recombinase; D019426:Integrases