Kim U J, Shizuya H, Chen X N, Deaven L, Speicher S, Solomon J, Korenberg J, Simon M I
Division of Biology and Beckmann Institute, California Institute of Technology, Pasadena 91125, USA.
Genet Anal. 1995 Oct;12(2):73-9. doi: 10.1016/1050-3862(95)00121-2.
Selection of chromosomal sublibraries from total human genomic libraries is critical for chromosome-based physical mapping approaches. We have previously reported a method of screening total human genomic library using flow sorted chromosomal DNA as a hybridization probe and selection of a human chromosome 22-enriched sublibrary from a total human bacterial artificial chromosome (BAC) library (Nucleic Acids Res 1995; 23: 1838-39). We describe here further details of the method of construction as well as characterization of the chromosome 22-enriched sublibrary thus constructed. Nearly 40% of the BAC clones that have been mapped by fluorescence in situ hybridization (FISH) analysis were localized to chromosome 22. By screening the sublibrary using chromosome 22-specific hybridization probes, we estimated that the sublibrary represents at least 2.5 x coverage of chromosome 22. This is in good agreement with the results from FISH mapping experiments. FISH map data also indicate that chromosome 22-specific BACs in the sublibrary represent all the subregions of chromosome 22.
从全人类基因组文库中挑选染色体亚文库对于基于染色体的物理图谱绘制方法至关重要。我们之前报道了一种使用流式分选的染色体DNA作为杂交探针筛选全人类基因组文库的方法，并从全人类细菌人工染色体（BAC）文库中挑选出了富含人类22号染色体的亚文库（《核酸研究》1995年；23: 1838 - 39）。我们在此描述构建该方法的更多细节以及由此构建的富含22号染色体的亚文库的特征。通过荧光原位杂交（FISH）分析定位的BAC克隆中，近40%定位于22号染色体。通过使用22号染色体特异性杂交探针筛选该亚文库，我们估计该亚文库至少覆盖22号染色体2.5倍。这与FISH图谱绘制实验的结果高度一致。FISH图谱数据还表明，该亚文库中22号染色体特异性BAC代表了22号染色体的所有亚区域。
