P[acman]:: A BAC transgenic platform for targeted insertion of large DNA fragments in D-melanogaster

被引：594

作者：

Venken, Koen J. T.

He, Yuchun

Hoskins, Roger A.

Bellen, Hugo J. ^{[1
]}

机构：

[1] Baylor Coll Med, Program Dev Biol, Houston, TX 77030 USA

[2] Baylor Coll Med, Dept Mol & Human Genet, Houston, TX 77030 USA

[3] Baylor Coll Med, Howard Hughes Med Inst, Houston, TX 77030 USA

[4] Univ Calif Berkeley, Lawrence Berkeley Lab, Dept Genome Biol, Berkeley, CA 94720 USA

[5] Baylor Coll Med, Dept Neurosci, Houston, TX 77030 USA

来源：

SCIENCE | 2006年 / 314卷 / 5806期

关键词：

D O I：

10.1126/science.1134426

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：1747 / 1751

页数：5

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