title: Simultaneous excision of two transgene flanking sequences and resolution of complex integration loci.
abstract: In planta excision techniques have proven useful both for basic biology and applied biotechnology. In this report, we describe a simple site-specific recombination (SSR) strategy that simultaneously removes pre-defined DNA sequences from both sides of a transgenic "gene of interest," leaving only the desired gene and short sequences from the recombinase recognition site. We have used the FLP/FRT SSR system to provide a proof of concept, though any of several other SSR systems could be used in the same way. The frequency of double excision ranged from 33% to 83% in different transgenic lines. We show that a single SSR reaction can simultaneously carry out double excisions and resolve complex transgene loci at high frequency. The method has direct biotechnological application and provides a useful tool for basic research.
authors.affiliation: Plant Gene Expression Lab, Plant Biology Department, NC State University, Campus Box 7550, Raleigh, NC 27695, USA. skumar3@unity.ncsu.edu
authors.forename: Sandeep
authors.initials: S
authors.lastname: Kumar
authors.forename: William F
authors.initials: WF
authors.lastname: Thompson
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D030821:Plants, Genetically Modified; D011995:Recombination, Genetic; D019076:Transgenes