Department of Molecular and Human Genetics, Howard Hughes Medical Institute, Baylor College of Medicine, Houston, Texas 77030, USA.
Genetics. 2010 Dec;186(4):1111-25. doi: 10.1534/genetics.110.121285. Epub 2010 Sep 27.
We describe a molecularly defined duplication kit for the X chromosome of Drosophila melanogaster. A set of 408 overlapping P[acman] BAC clones was used to create small duplications (average length 88 kb) covering the 22-Mb sequenced portion of the chromosome. The BAC clones were inserted into an attP docking site on chromosome 3L using ΦC31 integrase, allowing direct comparison of different transgenes. The insertions complement 92% of the essential and viable mutations and deletions tested, demonstrating that almost all Drosophila genes are compact and that the current annotations of the genome are reasonably accurate. Moreover, almost all genes are tolerated at twice the normal dosage. Finally, we more precisely mapped two regions at which duplications cause diplo-lethality in males. This collection comprises the first molecularly defined duplication set to cover a whole chromosome in a multicellular organism. The work presented removes a long-standing barrier to genetic analysis of the Drosophila X chromosome, will greatly facilitate functional assays of X-linked genes in vivo, and provides a model for functional analyses of entire chromosomes in other species.
我们描述了一个用于黑腹果蝇 X 染色体的分子定义的重复试剂盒。使用一组 408 个重叠的 P[acman] BAC 克隆,创建了覆盖染色体上已测序的 22-Mb 部分的小重复(平均长度为 88kb)。BAC 克隆使用 ΦC31 整合酶插入到染色体 3L 上的 attP 对接位点,允许直接比较不同的转基因。插入物补充了测试的必需和存活突变和缺失的 92%,表明几乎所有的果蝇基因都是紧凑的,并且当前的基因组注释相当准确。此外,几乎所有的基因都可以耐受两倍于正常剂量。最后,我们更精确地定位了两个导致雄性中重复引起二倍致死的区域。该集合包括第一个分子定义的重复试剂盒,用于覆盖多细胞生物中的整个染色体。所呈现的工作消除了黑腹果蝇 X 染色体遗传分析的长期障碍,将极大地促进体内 X 连锁基因的功能测定,并为其他物种的整个染色体的功能分析提供了模型。