Department of Biology, University of Szeged, H-6720 Szeged, Hungary.
G3 (Bethesda). 2012 Sep;2(9):1095-102. doi: 10.1534/g3.112.002899. Epub 2012 Sep 1.
Loss of the Y chromosome in Drosophila has no impact on cell viability and therefore allows us to assay the impact of environmental agents and genetic alterations on chromosomal loss. To detect in vivo chromosome loss in cells of the developing Drosophila wing primordia, we first engineered a Y chromosome with an attP docking site. By making use of the ΦC31 integrase system, we site-specifically integrated a genomic transgene encompassing the multiple wing hair (mwh) locus into this attP site, leading to a mwh(+)Y chromosome. This chromosome fully rescues the mwh mutant phenotype, an excellent recessive wing cell marker mutation. Loss of this mwh(+)Y chromosome in wing primordial cells then leads to manifestation of the mwh mutant phenotype in mwh-homozygous cells. The forming mwh clones permit us to quantify the effect of agents and genetic alterations by assaying frequency and size of the mwh mosaic spots. To illustrate the use of the mwh(+)Y loss system, the effects of four known mutagens (X-rays, colchicine, ethyl methanesulfonate, and formaldehyde) and two genetic conditions (loss- and gain-of-function lodestar mutant alleles) are documented. The procedure is simple, sensitive, and inexpensive.
Y 染色体的缺失对果蝇细胞的存活没有影响，因此我们可以检测环境因素和遗传改变对染色体缺失的影响。为了检测发育中的果蝇翅原基细胞中体内的染色体缺失，我们首先设计了一个带有 attP docking 位点的 Y 染色体。通过利用 ΦC31 整合酶系统，我们将一个包含多个翅毛（mwh）基因座的基因组转基因特异地整合到这个 attP 位点，导致 mwh(+)Y 染色体。这个染色体完全挽救了 mwh 突变体表型，这是一个极好的隐性翅膀细胞标记突变。mwh(+)Y 染色体在翅膀原基细胞中的缺失导致 mwh 纯合细胞中 mwh 突变体表型的表现。正在形成的 mwh 克隆使我们能够通过检测 mwh 镶嵌斑的频率和大小来量化试剂和遗传改变的影响。为了说明 mwh(+)Y 缺失系统的用途，记录了四种已知诱变剂（X 射线、秋水仙素、乙基甲磺酸酯和甲醛）和两种遗传条件（lodestar 突变等位基因的功能丧失和获得）的影响。该程序简单、灵敏、廉价。
