Golic K G, Golic M M
Department of Biology, University of Utah, Salt Lake City 84112, USA.
Genetics. 1996 Dec;144(4):1693-711. doi: 10.1093/genetics/144.4.1693.
We show that site-specific recombination can be used to engineer chromosome rearrangements in Drosophila melanogaster. The FLP site-specific recombinase acts on chromosomal target sites located within specially constructed P elements to provide an easy screen for the recovery of rearrangements with breakpoints that can be chosen in advance. Paracentric and pericentric inversions are easily recovered when two elements lie in the same chromosome in opposite orientation. These inversions are readily reversible. Duplications and deficiencies can be recovered by recombination between two elements that lie in the same orientation on the same chromosome or on homologues. We observe that the frequency of recombination between FRTs at ectopic locations decreases as the distance that separates those FRTs increases. We also describe methods to determine the absolute orientation of these P elements within the chromosome. The ability to produce chromosome rearrangements precisely between preselected sites provides a powerful new tool for investigations into the relationships between chromosome arrangement, structure, and function.
我们证明位点特异性重组可用于构建黑腹果蝇的染色体重排。FLP位点特异性重组酶作用于位于特殊构建的P元件内的染色体靶位点,为回收具有可预先选择的断点的重排提供了一个简单的筛选方法。当两个元件以相反方向位于同一条染色体上时,臂内倒位和臂间倒位很容易回收。这些倒位很容易逆转。重复和缺失可通过位于同一条染色体或同源染色体上相同方向的两个元件之间的重组来回收。我们观察到,异位位置的FRT之间的重组频率随着这些FRT之间距离的增加而降低。我们还描述了确定这些P元件在染色体内的绝对方向的方法。能够在预选位点之间精确产生染色体重排为研究染色体排列、结构和功能之间的关系提供了一个强大的新工具。