GENOMIC DELETION AND DUPLICATION COVERAGE IN DROSOPHILA

描述（由申请人提供）： 果蝇的功能基因组研究继续提供重要的发现，了解人类发展和改善人类健康。 发展研究工具，提高果蝇作为模式生物的效用，将有利于生物医学科学。 虽然果蝇的基因组现在已经测序，但果蝇遗传学家们仍然可以获得完整的染色体缺失和复制阵列，这将使他们能够研究所有染色体区域的基因功能。 作为对研究界的一项服务，BDSC的科学家们开始了一个项目，以提高删除和重复的覆盖率。 为了从开发的技术和在先前的资助下构建的遗传菌株中获益，他们建议：1）继续筛选染色体缺失，以提供基本上完全的常染色体缺失覆盖; 2）筛选染色体缺失，以提供类似的X染色体完全缺失覆盖;和3）产生X染色体区段的Y-连锁重复以提供X染色体的完全重复覆盖。 通过筛选涉及由P转座酶催化的P转座因子插入对的染色体重排，将分离提供新的基因组覆盖的细胞学大缺失。 在过去阻碍缺失恢复的单倍致死和单倍不育基因座将被缺失侧接以最大化基因组缺失覆盖。重复将分两步产生：使用FLP-FRT技术产生具有共同远端尖端断点和多个近端断点的X倒位，并附加到Y染色体上。 将对这些“X倒位+ Y”染色体进行辐照，以恢复从接近X末端倒位断点的可变位点延伸到X异染色质内的大的内部缺失。 所得的“X尖端-可变重复的X片段-X异染色质-Y”染色体将形成整个X染色体的Y连锁X重复的多个嵌套组。 在这个项目中开发的菌株将由BDSC分发，在那里它们将得到立即和广泛的使用。

DESCRIPTION (provided by applicant): Functional genomic research in Drosophila melanogaster continues to provide discoveries important to the understanding of human development and the improvement of human health. The development of research tools that enhance the utility of Drosophila as a model organism will benefit the biomedical sciences in general. Although the Drosophila genome is now sequenced, fly geneticists tack complete arrays of chromosomal deletions and duplications, which would enable them to study the functions of genes in all chromosomal regions. As a service to the research community, scientists at the BDSC began a project to improve deletion and duplication coverage. To benefit from techniques developed and genetic strains constructed under a previous grant, they propose to: 1) continue screening for chromosomal deletions to provide essentially complete deletion coverage of the autosomes; 2) screen for chromosomal deletions to provide similarly complete deletion coverage of the X chromosome; and 3) to generate Y-linked duplications of X chromosomal segments to provide complete duplication coverage of the X chromosome. Cytologically large deletions providing novel genome coverage will be isolated by screening for chromosomal rearrangements involving pairs of P transposable element insertions catalyzed by P transposase. Haplolethal and haplosterile loci that have hindered the recovery of deletions in the past will be flanked by deletions to maximize genomic deletion coverage. Duplications will be generated in two steps: X inversions with a common distal tip breakpoint and multiple proximal breakpoints will be made using FLP-FRT technology and appended to Y chromosomes. These "X inversion + Y" chromosomes will be irradiated to recover large internal deletions extending from variable sites proximal to the X tip inversion breakpoint to within X heterochromatin. The resulting "X tip-variable duplicated X segment-X heterochromatin-Y" chromosomes will form multiple nested sets of Y-linked X duplications for the entire X chromosome. Strains developed in this project will be distributed by the BDSC, where they will receive immediate and widespread use.