title: A recombineering-based gene tagging system for Arabidopsis.
abstract: Many of the experimental approaches aimed at studying gene function heavily rely on the ability to make precise modifications in the gene's DNA sequence. Homologous recombination (HR)-based strategies provide a convenient way to create such types of modifications. HR-based DNA sequence manipulations can be enormously facilitated by expressing in E. coli a small set of bacteriophage proteins that make the exchange of DNA between a linear donor and the target DNA molecules extremely efficient. These in vivo recombineering techniques have been incorporated as essential components of the molecular toolbox in many model organisms. In this chapter, we describe the experimental procedures involved in recombineering-based tagging of an Arabidopsis gene contained in a plant transformation-ready bacterial artificial chromosome (TAC).
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, 4558A Thomas Hall, Box 7612, Raleigh, NC, 27695-7612, USA, jmalonso@ncsu.edu.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Alonso
authors.forename: Anna N
authors.initials: AN
authors.lastname: Stepanova
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics* / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial / Q000737:chemistry / Q000378:metabolism*; D003001:Cloning, Molecular; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism*; D059765:Homologous Recombination; D016296:Mutagenesis