title: A simple polymerase chain reaction-based method for the construction of recombinase-mediated cassette exchange donor vectors.
abstract: Here we describe a simple method for generating donor vectors suitable for targeted transgenesis via recombinase-mediated cassette exchange (RMCE) using the PhiC31 integrase. This PCR-based strategy employs small attB "tails" on the primers used to amplify a sequence of interest, permitting the rapid creation of transgenes for in vivo analysis.
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Jack R
authors.initials: JR
authors.lastname: Bateman
authors.forename: C-ting
authors.initials: CT
authors.lastname: Wu
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D029721:Drosophila Proteins / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics*; D005260:Female; D018390:Gene Targeting; D005822:Genetic Vectors; D019426:Integrases / Q000378:metabolism; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D045522:Recombinases / Q000502:physiology*; D019076:Transgenes / Q000502:physiology*
chemical_list: D029721:Drosophila Proteins; D045522:Recombinases; D019426:Integrases
references.citation: Genetics. 2006 Jun;173(2):769-77
references.pmid: 16547094
references.citation: Biotechniques. 1990 May;8(5):528-35
references.pmid: 2357375
references.citation: Development. 2007 Oct;134(20):3571-84
references.pmid: 17905790
references.citation: Biochemistry. 1998 May 5;37(18):6229-34
references.pmid: 9572836
references.citation: Curr Opin Biotechnol. 2007 Oct;18(5):411-9
references.pmid: 17904350
references.citation: Science. 1982 Oct 22;218(4570):341-7
references.pmid: 6289435
references.citation: Dev Cell. 2004 Jan;6(1):7-28
references.pmid: 14723844
references.citation: Proc Natl Acad Sci U S A. 2000 May 23;97(11):5995-6000
references.pmid: 10801973
references.citation: Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3312-7
references.pmid: 17360644
references.citation: J Mol Biol. 1999 Oct 1;292(4):779-85
references.pmid: 10525404
references.citation: Curr Biol. 2002 Sep 17;12(18):1547-56
references.pmid: 12372246
references.citation: Science. 2006 Dec 15;314(5806):1747-51
references.pmid: 17138868
references.citation: Genetics. 2004 Apr;166(4):1775-82
references.pmid: 15126397