Golic M M, Rong Y S, Petersen R B, Lindquist S L, Golic K G
Department of Biology, University of Utah, Salt Lake City, UT 84112, USA.
Nucleic Acids Res. 1997 Sep 15;25(18):3665-71. doi: 10.1093/nar/25.18.3665.
The ability to place a series of gene constructs at a specific site in the genome opens new possibilities for the experimental examination of gene expression and chromosomal position effects. We report that the FLP- FRT site-specific recombination system of the yeast 2mu plasmid can be used to integrate DNA at a chromosomal FRT target site in Drosophila. The technique we used was to first integrate an FRT- flanked gene by standard P element-mediated transformation. FLP was then used to excise the FRT- flanked donor DNA and screen for FLP-mediated re-integration at an FRT target at a different chromosome location. Such events were recovered from up to 5% of the crosses used to screen for mobilization and are easily detectable by altered linkage of a white reporter gene or by the generation of a white + gene upon integration.
能够将一系列基因构建体置于基因组中的特定位点，为基因表达和染色体位置效应的实验研究开辟了新的可能性。我们报告称，酵母2μm质粒的FLP - FRT位点特异性重组系统可用于在果蝇的染色体FRT靶位点整合DNA。我们使用的技术是首先通过标准的P元件介导的转化整合一个侧翼带有FRT的基因。然后使用FLP切除侧翼带有FRT的供体DNA，并筛选在不同染色体位置的FRT靶位点处由FLP介导的重新整合。在用于筛选动员的杂交实验中，高达5%的杂交后代出现了此类事件，通过白色报告基因的连锁改变或整合时产生白色+基因很容易检测到。
