Golic K G
Howard Hughes Medical Institute, University of Chicago, Illinois 60637.
Genetics. 1994 Jun;137(2):551-63. doi: 10.1093/genetics/137.2.551.
The transposase source delta 2-3(99B) was used to mobilize a P element located at sites on chromosomes X, 2 and 3. The transposition event most frequently recovered was a chromosome with two copies of the P element at or near the original site of insertion. These were easily recognized because the P element carried a hypomorphic white gene with a dosage dependent phenotype; flies with two copies of the gene have darker eyes than flies with one copy. The P element also carried direct repeats of the recombination target (FRT) for the FLP site-specific recombinase. The synthesis of FLP in these flies caused excision of the FRT-flanked white gene. Because the two white copies excised independently, patches of eye tissue with different levels of pigmentation were produced. Thus, the presence of two copies of the FRT-flanked white gene could be verified. When the P elements lay in the same orientation, FLP-mediated recombination between the FRTs on separated elements produced deficiencies and duplications of the flanked region. When P elements were inverted, the predominant consequence of FLP-catalyzed recombination between the inverted elements was the formation of dicentric chromosomes and acentric fragments as a result of unequal sister chromatid exchange.
转座酶来源delta 2-3(99B)被用于激活位于X染色体、2号染色体和3号染色体上特定位置的P因子。最常检测到的转座事件是在原始插入位点或其附近带有两个P因子拷贝的染色体。这些很容易识别,因为P因子携带一个具有剂量依赖性表型的亚效白基因;携带两个该基因拷贝的果蝇眼睛比携带一个拷贝的果蝇眼睛颜色更深。P因子还携带FLP位点特异性重组酶的重组靶标(FRT)的同向重复序列。在这些果蝇中合成FLP会导致FRT侧翼的白基因切除。由于两个白基因拷贝独立切除,因此产生了色素沉着水平不同的眼组织斑块。因此,可以验证FRT侧翼白基因两个拷贝的存在。当P因子以相同方向排列时,FLP介导的分离元件上FRT之间的重组会产生侧翼区域的缺失和重复。当P因子反向排列时,FLP催化的反向元件之间的重组的主要结果是由于不等姐妹染色单体交换而形成双着丝粒染色体和无着丝粒片段。