大鼠高重复序列Rat(Hind Ⅲ)-1的染色体定位研究

The Studies on Localization of Rat Highly Repetitive Sequence Rat (Hind Ⅲ)-1 in the Chromosomes

以分离、纯化的370bp大小的Rat(HindⅢ)-1高重复序列为探针,用Biotin-Ⅱ-dUTP进行缺口翻译标记;与Wistar大鼠骨髓间期细胞和中期染色体进行原位杂交;用生物索化的碱性磷酸酶检测杂交结果。大鼠核型中除X,Y,1,2,12和20号染色体外,其余染色体着丝粒处均有杂交颗粒,以10,5,7,11,13号染色体所占比例较多。间期细胞核中亦可见到排列不规则的杂交颗粒。核杂交效率高于染色体杂交效率。Rat(HindⅢ)-1可能是Wistar大鼠特有的、普遍分布于染色体着丝粒处的一类高重复序列。

A highly repetitive sequence, Rat (Hind Ⅲ )-1 of 370 bp, was chosen to determine its chromosomal localiztion. Prepared firstly from the recombinant plasmid, and labeled with biotin-ll-dUTP by means of nick translation, biotiaylated probe was hybridized in situ to the interphase nuclei and the metaphase chromosomes of the bone marrow cell of Wistar rat. Finally, the Biotin-polyAP system was employed to detect the hybridizing sites. The results exhibited that hybridization grains, on the chromosomal centromeres of rat Karyotype, except X, Y, 1, 2, 12, 20 chromosomes, can be generally seen. The percentage of 10, 5, 7, 11, 13, 14 chromosomes among them were more higher. In the interphase nuclei, hybridizing grains distributed orderlessly. The statistical significance was appeared that the efficiency of nucleic hybridization was higher than that of chromosomal hybridization. Rat (HindⅢ )-l was considered as the Wistar rat-specific very basic unit of a highly repetitive sequence which distributed over chromosomal centromeres.