Broccoli D, Trevor K T, Miller O J, Miller D A
Department of Molecular Biology and Genetics, Wayne State University, Detroit, Michigan 48201.
Genomics. 1991 May;10(1):68-74. doi: 10.1016/0888-7543(91)90485-w.
Two cosmids (HRS-1 and HRS-2) containing mouse minor satellite DNA sequences have been isolated from a mouse genomic library. In situ hybridization under moderate stringency conditions to metaphase chromosomes from RCS-5, a tumor cell line derived from the SJL strain, mapped both HRS-1 and HRS-2 to the centromeric region of chromosome 4. Sequence data indicate that these cloned minor satellite DNA sequences have a basic higher order repeat of 180 bp, composed of three diverged 60-bp monomers. Digestion of mouse genomic DNA with several restriction enzymes produces a ladder of minor satellite fragments based on a 120-bp repeat. The restriction enzyme NlaIII (CATG) digests all the minor satellite DNA into three prominent bands of 120, 240, and 360 bp and a weak band of 180 bp. Thus, the majority of minor satellite sequences in the genome are arranged in repeats based on a 120-bp dimer, while the family of minor satellite sequences described here represents a rare variant of these sequences. Our results raise the possibility that there may be other variant families of minor satellites analogous to those of alphoid DNA present in humans.
从一个小鼠基因组文库中分离出了两个含有小鼠小卫星DNA序列的黏粒(HRS - 1和HRS - 2)。在适度严格条件下,对源自SJL品系的肿瘤细胞系RCS - 5的中期染色体进行原位杂交,将HRS - 1和HRS - 2都定位到了4号染色体的着丝粒区域。序列数据表明,这些克隆的小卫星DNA序列具有180 bp的基本高阶重复序列,由三个分化的60 bp单体组成。用几种限制酶消化小鼠基因组DNA会产生基于120 bp重复序列的小卫星片段阶梯。限制酶NlaIII(CATG)将所有小卫星DNA消化成120、240和360 bp的三条明显条带以及180 bp的一条弱条带。因此,基因组中的大多数小卫星序列以基于120 bp二聚体的重复形式排列,而这里描述的小卫星序列家族代表了这些序列的一种罕见变体。我们的结果提出了一种可能性,即可能存在其他类似于人类中存在的α卫星DNA的小卫星变体家族。