Qin S, Zhang J, Isaacs C M, Nagafuchi S, Jani Sait S N, Abel K J, Higgins M J, Nowak N J, Shows T B
Department of Human Genetics, Roswell Park Cancer Institute, New York State Department of Health, Buffalo 14263.
Genomics. 1993 Jun;16(3):580-5. doi: 10.1006/geno.1993.1233.
A targeted yeast artificial chromosome (YAC) library for chromosome 11 has been constructed from the J1 cell line that carries a single human chromosome 11 within a hamster DNA background. Interspecies chimeric clones generated during construction of the library were detected during the screening process and eliminated from the library. Contig assembly becomes much less difficult using such a library as the complexity is decreased and the ends of the clone inserts can be rescued for walking to neighboring clones. The library contains > 1824 clones with an average insert length of 337 kb. This represents a fourfold coverage of chromosome 11 or a > 95% chance of recovering a unique single-copy sequence from the library. Two hundred YAC clones were localized by fluorescence in situ hybridization and found to be randomly distributed along the chromosome. The library has been screened with probes for the chromosome 11 markers HBB, GLUR4, H19, and D11S193. Corresponding YAC clones have been isolated for each locus. This analysis has indicated that the library is unbiased, that cognate YAC clones can be recovered with chromosome 11 markers, and that extensive contig assembly should be feasible.
已从J1细胞系构建了一个针对11号染色体的酵母人工染色体（YAC）文库，该细胞系在仓鼠DNA背景中携带一条单一的人类11号染色体。在文库构建过程中产生的种间嵌合克隆在筛选过程中被检测到并从文库中剔除。使用这样的文库进行重叠群组装变得容易得多，因为复杂性降低了，并且克隆插入片段的末端可以被拯救出来以便向相邻克隆步移。该文库包含超过1824个克隆，平均插入长度为337 kb。这代表了11号染色体的四倍覆盖率，或者从文库中获得独特单拷贝序列的概率大于95%。通过荧光原位杂交对200个YAC克隆进行了定位，发现它们沿着染色体随机分布。用11号染色体标记HBB、GLUR4、H19和D11S193的探针筛选了该文库。已为每个位点分离出相应的YAC克隆。该分析表明该文库没有偏差，可以用11号染色体标记回收同源YAC克隆，并且广泛的重叠群组装应该是可行的。
