title: Genome manipulations with bacterial recombineering and site-specific integration in Drosophila.
abstract: Gene targeting is a vital tool for modern biology. The ability to efficiently and repeatedly target the same locus is made more efficient by the site-specific integrase mediated repeated targeting (SIRT) method, which combines homologous recombination, site-specific integration, and bacterial recombineering to conduct targeted modifications of individual loci. Here we describe the recombineering designs and procedures for the introduction of epitope tags, in-frame deletion mutations, and point mutations into plasmids that can later be used for SIRT.
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, National Cancer Institute (NCI), NIH, Bethesda, MD, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.forename: William
authors.initials: W
authors.lastname: Schreiner
authors.forename: Yikang S
authors.initials: YS
authors.lastname: Rong
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics; D001419:Bacteria / Q000235:genetics*; D004330:Drosophila / Q000235:genetics*; D018390:Gene Targeting; D005818:Genetic Engineering; D059765:Homologous Recombination; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010957:Plasmids / Q000235:genetics