GENOMIC DELETION AND DUPLICATION COVERAGE IN DROSOPHILA

描述(由申请人提供)： 对黑腹果蝇的功能基因组研究继续提供对理解人类发育和改善人类健康至关重要的发现。研究工具的开发将增强果蝇作为模式生物的实用性，这将使生物医学科学总体受益。尽管果蝇的基因组现在已经测序，但果蝇遗传学家利用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，以提高缺失和复制的覆盖率。为了从开发的技术和根据先前的资助构建的遗传菌株中受益，他们建议：1)继续筛查染色体缺失，以提供常染色体的基本完全缺失覆盖；2)染色体缺失筛查，以提供类似的X染色体的完全缺失覆盖；以及3)生成X染色体片段的Y连锁重复，以提供X染色体的完全复制覆盖。细胞学上提供新的基因组覆盖的大的缺失将通过筛选涉及由P转座酶催化的一对P转座元件插入的染色体重排来分离。过去阻碍缺失恢复的单致性和单倍性基因座将伴随着缺失，以最大限度地扩大基因组缺失的覆盖率。复制将分两步产生：使用FLP-FRT技术制作具有共同末端断裂点和多个近端断裂点的X倒位，并将其附加到Y染色体上。这些“X倒位+Y”染色体将被照射，以恢复从X末端倒位断裂点附近的可变点延伸到X异染色质内的大量内部缺失。由此产生的“X末端可变重复的X片段-X异染色质-Y”染色体将为整个X染色体形成多套Y连锁的X复制。该项目中开发的菌株将由BDSC分发，在那里它们将立即得到广泛使用。

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.