Charlieu J P, Laurent A M, Orti R, Viegas-Péquignot E, Bellis M, Roizès G
CNRS UPR 9008, Institut de Biologie, Montpellier, France.
Genomics. 1993 Mar;15(3):576-81. doi: 10.1006/geno.1993.1110.
A YAC clone from a chromosome 21-specific partial library was localized by in situ hybridization to the pericentromeric region of chromosomes 13 and 21 and to the long arm of chromosome 2, where an ancestral inactive centromere is present. Restriction mapping of the insert showed that it may contain tandemly repeated DNA. Probes for alpha-satellite and satellite II and III failed to hybridize with the cloned DNA. Shotgun subcloning might reveal a sequence that seems to be specific for chromosome 21. Alu-PCR was performed to generate probes from the YAC clone to map it more precisely, using a somatic hybrid containing only human chromosome 21. The inter-Alu sequences thus isolated were found to be clustered in an approximately 37-kb-long fragment common to chromosome 2, 13, and 21, which might be involved in the centromeric function of these chromosomes.
从21号染色体特异性部分文库中获得的一个酵母人工染色体(YAC)克隆,通过原位杂交定位到13号和21号染色体的着丝粒周围区域以及2号染色体的长臂,2号染色体长臂存在一个祖先非活性着丝粒。对插入片段进行限制性酶切图谱分析表明,它可能包含串联重复DNA。α-卫星以及卫星II和III的探针未能与克隆的DNA杂交。鸟枪法亚克隆可能会揭示一个似乎是21号染色体特有的序列。利用仅含人类21号染色体的体细胞杂种进行Alu-PCR,从YAC克隆中生成探针以更精确地对其进行定位。由此分离出的Alu间序列被发现聚集在一个约37kb长的片段中,该片段是2号、13号和21号染色体共有的,可能参与这些染色体的着丝粒功能。