Kuspa A, Vollrath D, Cheng Y, Kaiser D
Department of Biochemistry, Stanford University School of Medicine, CA 94305.
Proc Natl Acad Sci U S A. 1989 Nov;86(22):8917-21. doi: 10.1073/pnas.86.22.8917.
Random segments of Myxococcus xanthus DNA were cloned in yeast artificial chromosomes (YACs) to construct a physical map of the genome. EcoRI restriction maps of 409 YAC clones with inserts averaging 111 kilobase pairs (kb) were determined. Comparison to the map of a 300-kb region of M. xanthus obtained from clones in Escherichia coli indicates that segments of DNA cloned in YACs are stably maintained in yeast and that their sequences accurately reflect the structure of the Myxococcus genome. The 409 YAC inserts were ordered within 60 map segments (contigs) by aligning their EcoRI restriction maps and by hybridization with 18 gene-specific DNA probes. These 60 map segments may represent the entire Myxococcus genome and could be used to organize its genetic information. This study illustrates the utility of YACs for cloning large segments of DNA and for reliable long-range genomic mapping.
将黄色粘球菌（Myxococcus xanthus）DNA的随机片段克隆到酵母人工染色体（YAC）中，以构建基因组物理图谱。测定了409个YAC克隆的EcoRI限制性图谱，这些克隆的插入片段平均为111千碱基对（kb）。与从大肠杆菌克隆获得的黄色粘球菌300 kb区域的图谱比较表明，克隆到YAC中的DNA片段在酵母中能稳定保存，且其序列准确反映了黄色粘球菌基因组的结构。通过比对409个YAC插入片段的EcoRI限制性图谱，并与18个基因特异性DNA探针杂交，将这些插入片段排列在60个图谱片段（重叠群）中。这60个图谱片段可能代表整个黄色粘球菌基因组，可用于整理其遗传信息。本研究说明了YAC在克隆大片段DNA和进行可靠的长距离基因组作图方面的实用性。
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