Rocchi M, Archidiacono N, Ward D C, Baldini A
Laboratorio di Genetica Molecolare, Istituto G. Gaslini, Sassari, Italy.
Genomics. 1991 Mar;9(3):517-23. doi: 10.1016/0888-7543(91)90419-f.
We have isolated a DNA clone (pMR9A) that identifies an alphoid DNA subset specific for chromosome 9. This alphoid subset is characterized by a dimeric organization as revealed by Southern blot analysis after digestion with HaeIII, HinfI, or StuI. Nonradioactive in situ hybridization demonstrated that pMR9A hybridizes only to the centromeric region of chromosome 9 and reveals chromosome 9 aneuploidies in interphase nuclei. In addition, the probe detects quantitative differences in alpha satellite DNA on chromosome 9, but these quantitative differences are not correlated with the size of the heterochromatic region. Double-labeling experiments, using a chromosome 9-specific satellite 3 clone and pMR9A, enabled us spatially to distinguish the alphoid and satellite 3 domains on metaphase chromosomes after treatment of the cultures with 5-azacytidine.
我们分离出了一个DNA克隆（pMR9A），它可识别9号染色体特有的α卫星DNA亚群。经HaeIII、HinfI或StuI消化后的Southern印迹分析显示，该α卫星亚群具有二聚体结构。非放射性原位杂交表明，pMR9A仅与9号染色体的着丝粒区域杂交，并揭示了间期核中9号染色体的非整倍性。此外，该探针检测到9号染色体上α卫星DNA的数量差异，但这些数量差异与异染色质区域的大小无关。使用9号染色体特异性卫星3克隆和pMR9A进行的双标记实验，使我们能够在用5-氮杂胞苷处理培养物后，在中期染色体上从空间上区分α卫星和卫星3结构域。
