Auriche C, Donini P, Ascenzioni F
Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Biologia Cellulare e dello Sviluppo, University of Rome La Sapienza, Italy.
EMBO Rep. 2001 Feb;2(2):102-7. doi: 10.1093/embo-reports/kve018.
Mammalian artificial chromosomes (MACs) provide a new tool for the improvement of our knowledge of chromosome structure and function. Moreover, they constitute an alternative and potentially powerful tool for gene delivery both in cultured cells and for the production of transgenic animals. In the present work we describe the molecular structure of MC1, a human minichromosome derived from chromosome 1. By means of restriction and hybridization analysis, satellite-PCR, in situ hybridization on highly extended chromatin fibres, and indirect immunofluorescence, we have established that: (i) MC1 has a size of 5.5 Mb; (ii) it consists of 1.1 Mb alphoid, 3.5 Mb Sat2 DNA, and telomeric and subtelomeric sequences at both ends; (iii) it contains an unusual region of interspersed Sat2 and alphoid DNAs at the junction of the alphoid and the Sat2 blocks; and (iv) the two alphoid blocks and the Sat2-alphoid region bind centromeric proteins suggesting that they participate in the formation of a functional kinetochore.
哺乳动物人工染色体（MACs）为增进我们对染色体结构和功能的了解提供了一种新工具。此外，它们构成了一种用于在培养细胞中进行基因传递以及生产转基因动物的替代且潜在强大的工具。在本研究中，我们描述了源自1号染色体的人类小染色体MC1的分子结构。通过限制性内切酶和杂交分析、卫星PCR、在高度伸展的染色质纤维上进行原位杂交以及间接免疫荧光，我们确定了：（i）MC1大小为5.5 Mb；（ii）它由1.1 Mb的α卫星DNA、3.5 Mb的Sat2 DNA以及两端的端粒和亚端粒序列组成；（iii）在α卫星和Sat2区域的交界处包含一个由Sat2和α卫星DNA穿插组成的异常区域；（iv）两个α卫星区域以及Sat2-α卫星区域结合着着丝粒蛋白，表明它们参与了功能性动粒的形成。
