Stone J C, Dower N A, Hauseman J, Cseko Y M, Sederoff R
Can J Genet Cytol. 1983 Apr;25(2):129-38. doi: 10.1139/g83-025.
Using a "single-fly" nucleic acid hybridization method, we have surveyed a collection of D. melanogaster strains in search of variants which affect DNA complementary to the polypyrimidine sequence corresponding to one strand of the 1.705 satellite. Hybridization of labelled polypyrimidine probe to polypurine sequence in nucleic acid extracts of single flies, followed by thermal chromatography over hydroxyapatite led to the identification of one variant. The strain Cy/M(2)S2(10) produced excess hybrid, much of which had low thermal stability. A developmental analysis of the low-melt hybrid phenotype showed that certain tissues, in particular the ovaries, were affected. In addition to the biochemical phenotype, the break down of nurse cell nuclei in Cy/M(2)S2(10) ovaries during oocyte maturation was abnormal. A genetic analysis demonstrated that both the biochemical and cytological phenotypes were the consequences of a single recessive mutation in the DNase-1 gene on chromosome III. Studies with purified DNA demonstrated that the low-melt hybrid phenotype resulted from the accumulation of low molecular weight DNA complementary to the polypyrimidine probe.
我们采用“单蝇”核酸杂交方法，对一组黑腹果蝇品系进行了研究，以寻找影响与对应于1.705卫星一条链的多嘧啶序列互补的DNA的变体。将标记的多嘧啶探针与单蝇核酸提取物中的多嘌呤序列杂交，然后在羟基磷灰石上进行热色谱分析，从而鉴定出一个变体。品系Cy/M(2)S2(10)产生了过量的杂交体，其中大部分热稳定性较低。对低熔点杂交体表型的发育分析表明，某些组织，特别是卵巢受到了影响。除了生化表型外，Cy/M(2)S2(10)卵巢中卵母细胞成熟过程中滋养细胞核的解体也不正常。遗传分析表明，生化和细胞学表型都是Ⅲ号染色体上DNase-1基因单个隐性突变的结果。对纯化DNA的研究表明，低熔点杂交体表型是与多嘧啶探针互补的低分子量DNA积累的结果。
