Development of a more efficient insect transformation system

拟议研究的长期目标是开发 物理和遗传学系统，用于将克隆的DNA传递到任何 昆虫物种，随后位点特异性整合到 基因组 我们将探索黑腹果蝇 胚胎的卵黄膜已经被一种 最近开发的一种方法， 前体 我们还将利用果蝇胚胎， 两种主要脱氧核糖核酸酶的突变体，以增加 导入的DNA在体细胞和胚芽中的持久性 细胞 本研究的第三个目的是调整“翻转 用于昆虫宿主的来自酵母的“重组酶”系统。 最初 果蝇和家蝇（Musca arteritica）的胚胎将被 用重组位点序列（FRT）转化。 然后 重组酶基因将与选择标记一起沿着引入 含有单个FRT序列的质粒中的基因。 位点特异性 标记基因的重组将由基因组学监测。 Southern和染色体原位杂交分析。 因为 这样一种更有效和定点的转化系统， 是为果蝇开发的一个模型系统， 到任何其他昆虫物种，将有可能引入基因， 随意转化为害虫和人类疾病的载体。 这 生物学家应该有能力为昆虫设计新的策略 控制。*//

The long term objective of the proposed research is to develop physical and genetics systems for the delivery of cloned DNA to any insect species with subsequent site specific integration into the genome. We will explore the ability of Drosophila melanogaster embryos whose vitelline membranes have been permeabilized by a recently developed procedure to take up both DNA and nucleotide precursors. We will also utilize Drosophila embryos which are mutant for two predominant deoxyribonucleases to increase the persistence of the introduced DNA in both the somatic and germ cells. The third aim of the research is to adapt the "Flip recombinase" system from yeast for use in insect hosts. Initially Drosophila and Musca domestica (housefly) embryos will be transformed with the recombination site sequence (FRT). Then, the recombinase gene will be introduced along with selectable marker genes in plasmids containing a single FRT sequence. Site specific recombination of the marker genes will be monitored by genomic Southern and chromosomal in situ hybridization analysis. Because such a more efficient and site directed transformation system that is developed for Drosophila as a model system should be applicable to any other insect species, it will be possible to introduce genes at will into insect pests and vectors of human disease. This ability should allow biologists to devise new strategies for insect control.***//