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9号染色体短臂的辐射杂种与酵母人工染色体整合图谱。

Integrated radiation hybrid and yeast artificial chromosome map of chromosome 9p.

作者信息

Bouzyk M, Bryant S P, Evans C, Guioli S, Ford S, Schmidt K, Goodfellow P N, Povey S, Rebello M, Rousseaux S, Spurr N K

机构信息

Human Genetic Resources, Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, UK.

出版信息

Eur J Hum Genet. 1997 Sep-Oct;5(5):299-307.

PMID:9412787
Abstract

A panel of 93 radiation-reduced hybrids have been screened using PCR amplification and oligonucleotide primers for sequence-tagged sites (STSs) specific for 114 single-copy loci mapping to the short arm of chromosome 9. An x-ray dose of 6,000 rads gave an average retention frequency of approximately 23%. We have constructed a framework map containing 31 markers ordered by analyzing coretention patterns, with support for the order greater than 1,000:1. In addition, we have placed the remaining markers which could not be mapped to a single interval with this support to a range of intervals on the framework map. The STS oligonucleotide primers used in the construction of the radiation hybrid (RH) map have been used to isolate and order yeast artificial chromosomes (YACs) assigned to 9p identified from the CEPH megaYAC library. Eighty-nine STS markers have screened positive with at least one YAC. A total of 88 individual YACs (with an average size of 0.9 MB) have been placed on the map in a series of contigs and in some cases mapped cytogenetically by fluorescence in situ hybridization. Additionally, the YAC information has been used in conjunction with the RH framework placements to generate an integrated map containing 65 loci including 51 uniquely positioned markers, with an average resolution of 0.79 Mb.

摘要

利用聚合酶链反应(PCR)扩增和针对114个单拷贝基因座的序列标签位点(STS)的寡核苷酸引物,对一组93个辐射减少的杂种进行了筛选,这些基因座定位于9号染色体短臂。6000拉德的X射线剂量产生的平均保留频率约为23%。我们通过分析共保留模式构建了一个包含31个标记的框架图,对顺序的支持度大于1000:1。此外,我们已将在这种支持下无法定位到单个区间的其余标记放置在框架图上的一系列区间内。用于构建辐射杂种(RH)图谱的STS寡核苷酸引物已用于分离和排序从CEPH超级酵母人工染色体(YAC)文库中鉴定出的、分配到9p的酵母人工染色体(YAC)。89个STS标记已筛选出至少与一个YAC呈阳性反应。总共88个单独的YAC(平均大小为0.9兆碱基)已通过一系列重叠群放置在图谱上,并且在某些情况下通过荧光原位杂交进行了细胞遗传学定位。此外,YAC信息已与RH框架定位结合使用,以生成一个包含65个基因座的综合图谱,其中包括51个唯一定位的标记,平均分辨率为0.79兆碱基。

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