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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >End-sequence profiling: Sequence-based analysis of aberrant genomes
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End-sequence profiling: Sequence-based analysis of aberrant genomes

机译：末端序列分析：异常基因组的基于序列的分析

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摘要

Genome rearrangements are important in evolution, cancer, and other diseases. Precise mapping of the rearrangements is essential for identification of the involved genes, and many techniques have been developed for this purpose. We show here that end-sequence profiling (ESP) is particularly well suited to this purpose. ESP is accomplished by constructing a bacterial artificial chromosome (BAC) library from a test genome, measuring BAC end sequences, and mapping end-sequence pairs onto the normal genome sequence. Plots of BAC end-sequences density identify copy number abnormalities at high resolution. BACs spanning structural aberrations have end pairs that map abnormally far apart on the normal genome sequence. These pairs can then be sequenced to determine the involved genes and breakpoint sequences. ESP analysis of the breast cancer cell line MCF-7 demonstrated its utility for analysis of complex genomes. End sequencing of approximate to8,000 clones (0.37-fold haploid genome clonal coverage) produced a comprehensive genome copy number map of the MCF-7 genome at better than 300-kb resolution and identified 381 genome breakpoints, a subset of which was verified by fluorescence in situ hybridization mapping and sequencing. [References: 19]
机译：基因组重排在进化，癌症和其他疾病中很重要。重排的精确作图对于鉴定所涉及的基因至关重要，为此已经开发了许多技术。我们在这里表明，末端序列分析（ESP）特别适合于此目的。通过从测试基因组构建细菌人工染色体（BAC）文库，测量BAC末端序列，并将末端序列对映射到正常基因组序列，可以实现ESP。 BAC末端序列密度图可识别高分辨率下的拷贝数异常。跨越结构性畸变的BAC的末端对在正常基因组序列上的异常距离很远。然后可以对这些对进行测序，以确定涉及的基因和断点序列。乳腺癌细胞系MCF-7的ESP分析证明了其可用于复杂基因组分析。大约8,000个克隆的末端测序（0.37倍的单倍体基因组克隆覆盖率）产生了MCF-7基因组的全面基因组拷贝数图，分辨率超过300-kb，并鉴定了381个基因组断点，其中一个子集已通过验证荧光原位杂交作图和测序。 [参考：19]

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