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Originally published in Science Express on 30 November 2006
Science 15 December 2006:
Vol. 314. no. 5806, pp. 1747 - 1751
DOI: 10.1126/science.1134426
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Research Articles

P[acman]: A BAC Transgenic Platform for Targeted Insertion of Large DNA Fragments in D. melanogaster

Koen J. T. Venken,1 Yuchun He,2,3 Roger A. Hoskins,4 Hugo J. Bellen1,2,3,5*

We describe a transgenesis platform for Drosophila melanogaster that integrates three recently developed technologies: a conditionally amplifiable bacterial artificial chromosome (BAC), recombineering, and bacteriophage {Phi}C31–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. {Phi}C31-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.

1 Program in Developmental Biology, Baylor College of Medicine, Houston, TX 77030, USA.
2 Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA.
3 Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX 77030, USA.
4 Department of Genome Biology, Lawrence Berkeley National Laboratory, Berkeley, CA 94720–3200, USA.
5 Department of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA.

* To whom correspondence should be addressed. E-mail: hbellen{at}bcm.edu

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