P[acman]:: A BAC transgenic platform for targeted insertion of large DNA fragments in D-melanogaster
P[acman]:: A BAC transgenic platform for targeted insertion of large DNA fragments in D-melanogaster
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DOI：
10.1126/science.1134426
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发表时间：
2006-12-15
期刊：
影响因子：
56.9
通讯作者：
Bellen, Hugo J.
中科院分区：
文献类型：
--
作者：
Venken, Koen J. T.;He, Yuchun;Bellen, Hugo J.
我们描述了一个黑胃果蝇的转基因平台，该平台整合了三种最新开发的技术：条件扩增细菌人工染色体（BAC）、重组和噬菌体phi c31介导的转基因。BAC维持在低拷贝数，有利于质粒的维持和重组，但被诱导到高拷贝数，便于质粒的分离。重组允许细菌的间隙修复和诱变。间隙修复可以有效地从P1或BAC克隆中提取长达133千碱基的DNA片段。phi c31介导的转基因将这些大的DNA片段整合到基因组的特定位点，从而挽救相应基因的致命突变。这个转基因平台将极大地促进大多数果蝇基因的结构/功能分析。
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.