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首页> 外文期刊>Molecular and Cellular Biology >CpG island mapping of a mouse double-minute chromosome.
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CpG island mapping of a mouse double-minute chromosome.

机译：小鼠双倍染色体的CpG岛定位。

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The development of double-minute chromosomes (DMs) and subsequent gene amplification are important genomic alterations resulting in increased oncogene expression in a variety of tumors. The molecular mechanisms mediating the development of these acentric extrachromosomal elements have not been completely defined. To elucidate the mechanisms involved in DM formation, we have developed strategies to map amplified circular DM DNA. In this study, we present a long-range restriction map of a 980-kb DM. A cell line cloned from mouse EMT-6 cells was developed by stepwise selection for resistance to methotrexate. This cloned cell line contains multiple copies of the 980-kb DM carrying the dihydrofolate reductase (DHFR) gene. A long-range restriction map was developed in which a hypomethylated CpG-rich region near the DHFR gene served as a landmark. This strategy was combined with plasmid-like analysis of ethidium bromide-stained pulsed-field gels and indicated that a single copy of the DHFR gene was located near a hypomethylated region containing SsII and NotI sites. At least 490 kb of this DM appears to be composed of unrearranged chromosomal DNA.
机译：两分钟染色体（DMs）的发展和随后的基因扩增是重要的基因组改变，可导致多种肿瘤中癌基因表达的增加。介导这些中心染色体外元件发展的分子机制尚未完全确定。为了阐明参与DM形成的机制，我们已开发出策略来绘制扩增的环状DM DNA。在这项研究中，我们提出了980-kb DM的远距离限制图。通过逐步选择对甲氨蝶呤的抗性，开发了从小鼠EMT-6细胞克隆的细胞系。此克隆的细胞系包含带有二氢叶酸还原酶（DHFR）基因的980-kb DM的多个副本。制定了一个远程限制性图谱，其中DHFR基因附近的一个低甲基化的富含CpG的区域充当了标志。该策略与溴化乙锭染色的脉冲场凝胶的质粒样分析相结合，表明DHFR基因的单个拷贝位于包含SsII和NotI位点的低甲基化区域附近。至少490 kb的该DM似乎由未重排的染色体DNA组成。

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