Brody H, Griffith J, Cuticchia A J, Arnold J, Timberlake W E
Department of Genetics, University of Georgia, Athens 30602.
Nucleic Acids Res. 1991 Jun 11;19(11):3105-9. doi: 10.1093/nar/19.11.3105.
Development of physical genomic maps is facilitated by identification of overlapping recombinant DNA clones containing long chromosomal DNA inserts. To simplify the analysis required to determine which clones in a genomic library overlap one another, we partitioned Aspergillus nidulans cosmid libraries into chromosome-specific subcollections. The eight A. nidulans chromosomes were resolved by pulsed field gel electrophoresis and hybridized to filter replicas of cosmid libraries. The subcollections obtained appeared to be representative of the chromosomes based on the correspondence between subcollection size and chromosome length. A sufficient number of clones was obtained in each chromosome-specific subcollection to predict the overlap and assembly of individual clones into a limited number of contiguous regions. This approach should be applicable to many organisms whose genomes can be resolved by pulsed field gel electrophoresis.
通过鉴定包含长染色体DNA插入片段的重叠重组DNA克隆，有助于构建物理基因组图谱。为了简化确定基因组文库中哪些克隆相互重叠所需的分析，我们将构巢曲霉粘粒文库划分为染色体特异性亚文库。通过脉冲场凝胶电泳分离出构巢曲霉的八条染色体，并与粘粒文库的滤膜复制品杂交。根据亚文库大小与染色体长度之间的对应关系，获得的亚文库似乎代表了相应的染色体。在每个染色体特异性亚文库中获得了足够数量的克隆，以预测单个克隆的重叠情况并将其组装成有限数量的连续区域。这种方法应该适用于许多基因组可以通过脉冲场凝胶电泳解析的生物。
