A simple polymerase chain reaction-based method for the construction of recombinase-mediated cassette exchange donor vectors.
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.
Bateman
Jack R
JR
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Wu
C-ting
CT
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*
D029721:Drosophila Proteins; D045522:Recombinases; D019426:Integrases
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