O'Gorman S, Fox D T, Wahl G M
Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA 92037.
Science. 1991 Mar 15;251(4999):1351-5. doi: 10.1126/science.1900642.
A binary system for gene activation and site-specific integration, based on the conditional recombination of transfected sequences mediated by the FLP recombinase from yeast, was implemented in mammalian cells. In several cell lines, FLP rapidly and precisely recombined copies of its specific target sequence to activate an otherwise silent beta-galactosidase reporter gene. Clones of marked cells were generated by excisional recombination within a chromosomally integrated copy of the silent reporter. By the reverse reaction, integration of transfected DNA was targeted to a specific chromosomal site. The results suggest that FLP could be used to mosaically activate or inactivate transgenes for analysis of vertebrate development, and to efficiently integrate transfected DNA at predetermined chromosomal locations.
基于酵母FLP重组酶介导的转染序列的条件重组,构建了一种用于基因激活和位点特异性整合的二元系统,并在哺乳动物细胞中得以应用。在多种细胞系中,FLP能快速且精确地重组其特定靶序列的拷贝,从而激活原本沉默的β-半乳糖苷酶报告基因。通过在沉默报告基因的染色体整合拷贝内进行切除重组,产生了标记细胞的克隆。通过反向反应,可将转染DNA靶向整合到特定的染色体位点。这些结果表明,FLP可用于镶嵌式激活或失活转基因,以分析脊椎动物的发育,并能有效地将转染DNA整合到预定的染色体位置。