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P[acman]: A BAC Transgenic Platform for Targeted Insertion of Large DNA Fragments in D. melanogaster
Koen J. T. Venken, Yuchun He, Roger A. Hoskins and Hugo J. Bellen
New Series, Vol. 314, No. 5806 (Dec. 15, 2006), pp. 1747-1751
Published by: American Association for the Advancement of Science
Stable URL: http://www.jstor.org/stable/20035035
Page Count: 5
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We describe a transgenesis platform for Drosophila melanogaster that integrates three recently developed technologies: a conditionally amplifiable bacterial artificial chromosome (BAC), recombineering, and bacteriophage φC31--mediated transgenesis. The BAC is maintained at low copy number, facilitating plasmid maintenance and recombineering, but is induced to high copy number for plasmid isolation. Recombineering allows gap repair and mutagenesis in bacteria. Gap repair efficiently retrieves DNA fragments up to 133 kilobases long from P1 or BAC clones. φC31-mediated transgenesis integrates these large DNA fragments at specific sites in the genome, allowing the rescue of lethal mutations in the corresponding genes. This transgenesis platform should greatly facilitate structure/function analyses of most Drosophila genes.
Science © 2006 American Association for the Advancement of Science