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迈向人类10号染色体的物理图谱:183个代表80个基因座的酵母人工染色体的分离以及通过荧光原位杂交对94个酵母人工染色体进行区域定位

Toward a physical map of human chromosome 10: isolation of 183 YACs representing 80 loci and regional assignment of 94 YACs by fluorescence in situ hybridization.

作者信息

Moir D T, Dorman T E, Day J C, Ma N S, Wang M T, Mao J I

机构信息

Department of Human and Molecular Genetics, Collaborative Research, Inc., Waltham, Massachusetts 02154.

出版信息

Genomics. 1994 Jul 1;22(1):1-12. doi: 10.1006/geno.1994.1338.

Abstract

One hundred eighty-three YACs carrying human chromosome 10 sequences were isolated from multigenome equivalent libraries by PCR-based screening for the presence of 80 different chromosome 10-specific STSs. Ninety-four of the isolated YACs, representing 52 genes and DNA segments, were mapped to regions of chromosome 10 by fluorescence in situ hybridization. The results localized 26 DNA segments to cytogenetic bands for the first time. About 37% (35/94) of the YACs hybridized to more than one chromosomal location: 31 to other chromosomes in addition to chromosome 10 and 4 to 2 distinct locations on chromosome 10. These results are consistent with the number of chimeric YACs expected from these libraries but may also reflect the presence of 2 or more YACs within a single clone or the presence of low copy repeated elements within the genome. This STS anchor screening effort resulted in the identification of 69 contigs, with 7 contigs consisting of 2 anchors each and 1 contig consisting of 5 anchors. All linked STSs were multiply linked by at least 2 independent YACs. These anchored YACs span the entire chromosome and appear to cover 15% of chromosome 10.

摘要

通过基于聚合酶链反应(PCR)的筛选方法,从多基因组当量文库中分离出183个携带人类10号染色体序列的酵母人工染色体(YAC),以检测80种不同的10号染色体特异性序列标签位点(STS)的存在情况。通过荧光原位杂交,将分离得到的94个YAC(代表52个基因和DNA片段)定位到10号染色体区域。这些结果首次将26个DNA片段定位到细胞遗传学带。约37%(35/94)的YAC与不止一个染色体位置杂交:31个除了与10号染色体杂交外,还与其他染色体杂交,4个与10号染色体上2个不同位置杂交。这些结果与这些文库中预期的嵌合YAC数量一致,但也可能反映单个克隆中存在2个或更多YAC,或者基因组中存在低拷贝重复元件。这种STS锚定筛选工作导致鉴定出69个重叠群,其中7个重叠群各由2个锚定序列组成,1个重叠群由5个锚定序列组成。所有相连的STS都由至少2个独立的YAC多重连接。这些锚定的YAC跨越整个染色体,似乎覆盖了10号染色体的15%。

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