title: The pollen- and embryo-specific Arabidopsis DLL promoter bears good potential for application in marker-free Cre/loxP self-excision strategy.
abstract: Marker-free transgenic plants can be generated with high efficiency by using the Cre/ lox P self-excision system controlled by the pollen- and embryo-specific Arabidopsis DLL promoter. In this work, we aimed to study the feasibility of using the pollen- and embryo-specific DLL promoter of the At4g16160 gene from Arabidopsis thaliana in a Cre/loxP self-excision strategy. A Cre/loxP self-excision cassette controlled by the DLL promoter was introduced into the tobacco genome via Agrobacterium-mediated transformation. No evidence for premature activation of the Cre/loxP system was observed in primary transformants. The efficiency of nptII removal during pollen and embryo development was investigated in transgenic T1 progenies derived from eight self- and four cross-pollinated T0 lines, respectively. Segregation and rooting assays were performed to select recombined T1 plants. Molecular analyses of these plants confirmed the excision event in all analysed T0 lines and marker-free transgenic T1 plants were obtained with efficiency of up to 96.2%. The Arabidopsis DLL promoter appears to be a strong candidate to drive Cre-mediated recombination not only in tobacco as a model plant, but also in other plant species.
authors.affiliation: Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Akademicka 2, P.O. Box 39A, 95 007, Nitra, Slovak Republic, zuzana.poloniova@savba.sk.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Polóniová
authors.forename: Martin
authors.initials: M
authors.lastname: Jopčík
authors.forename: Ildikó
authors.initials: I
authors.lastname: Matušíková
authors.forename: Jana
authors.initials: J
authors.lastname: Libantová
authors.forename: Jana
authors.initials: J
authors.lastname: Moravčíková
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics*; D001483:Base Sequence; D018506:Gene Expression Regulation, Plant; D005819:Genetic Markers; D005822:Genetic Vectors; D019426:Integrases / Q000235:genetics; D007473:Ion Channels / Q000235:genetics*; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D011058:Pollen / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012639:Seeds / Q000235:genetics; D050610:Sodium-Phosphate Cotransporter Proteins, Type II / Q000235:genetics; D014026:Nicotiana / Q000235:genetics*; D014169:Transformation, Bacterial
chemical_list: D029681:Arabidopsis Proteins; D005819:Genetic Markers; D007473:Ion Channels; C512221:OEP16-L protein, Arabidopsis; D050610:Sodium-Phosphate Cotransporter Proteins, Type II; C045073:Cre recombinase; D019426:Integrases