Morris A C, Schaub T L, James A A
Department of Molecular Biology & Biochemistry, University of California, Irvine 92717.
Nucleic Acids Res. 1991 Nov 11;19(21):5895-900. doi: 10.1093/nar/19.21.5895.
The activity of a yeast recombinase, FLP, on specific target DNA sequences, FRT, has been demonstrated in embryos of the vector mosquito, Aedes aegypti. In a series of experiments, plasmids containing the FLP recombinase under control of a heterologous heat-shock gene promoter were co-injected with target plasmids containing FRT sites into preblastoderm stage mosquito embryos. FLP-mediated recombination was detected between (i) tandem repeats of FRT sites leading to the excision of specific DNA sequences and (ii) FRT sites located on separate plasmids resulting in the formation of heterodimeric or higher order multimeric plasmids. In addition to FRT sites originally isolated from the yeast 2 microns plasmid, a number of synthetic FRT sites were also used. The synthetic sites were fully functional as target sites for recombination and gave results similar to those derived from the yeast 2 microns plasmid. This successful demonstration of yeast FLP recombinase activity in the mosquito embryo suggests a possible future application of this system in establishing transformed lines of mosquitoes for use in vector control strategies and basic studies.
酵母重组酶FLP对特定靶DNA序列FRT的活性已在媒介蚊虫埃及伊蚊的胚胎中得到证实。在一系列实验中，将含有在异源热休克基因启动子控制下的FLP重组酶的质粒与含有FRT位点的靶质粒共同注射到胚盘形成前期的蚊虫胚胎中。在以下情况中检测到了FLP介导的重组：（i）FRT位点的串联重复导致特定DNA序列的切除；（ii）位于不同质粒上的FRT位点导致异二聚体质粒或更高阶多聚体质粒的形成。除了最初从酵母2μm质粒中分离出的FRT位点外，还使用了一些合成FRT位点。这些合成位点作为重组的靶位点具有完全功能，并且产生的结果与源自酵母2μm质粒的结果相似。酵母FLP重组酶活性在蚊虫胚胎中的这一成功证明表明，该系统未来可能应用于建立用于病媒控制策略和基础研究的转基因蚊虫品系。
