Lim A, Dimalanta E T, Potamousis K D, Yen G, Apodoca J, Tao C, Lin J, Qi R, Skiadas J, Ramanathan A, Perna N T, Plunkett G, Burland V, Mau B, Hackett J, Blattner F R, Anantharaman T S, Mishra B, Schwartz D C
Laboratory for Molecular and Computational Genomics, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Genome Res. 2001 Sep;11(9):1584-93. doi: 10.1101/gr.172101.
We have constructed NheI and XhoI optical maps of Escherichia coli O157:H7 solely from genomic DNA molecules to provide a uniquely valuable scaffold for contig closure and sequence validation. E. coli O157:H7 is a common pathogen found in contaminated food and water. Our approach obviated the need for the analysis of clones, PCR products, and hybridizations, because maps were constructed from ensembles of single DNA molecules. Shotgun sequencing of bacterial genomes remains labor-intensive, despite advances in sequencing technology. This is partly due to manual intervention required during the last stages of finishing. The applicability of optical mapping to this problem was enhanced by advances in machine vision techniques that improved mapping throughput and created a path to full automation of mapping. Comparisons were made between maps and sequence data that characterized sequence gaps and guided nascent assemblies.
我们仅从基因组DNA分子构建了大肠杆菌O157:H7的NheI和XhoI光学图谱,以提供一个对重叠群封闭和序列验证具有独特价值的支架。大肠杆菌O157:H7是在受污染的食物和水中发现的常见病原体。我们的方法无需分析克隆、PCR产物和杂交,因为图谱是由单个DNA分子的集合构建而成的。尽管测序技术有所进步,但细菌基因组的鸟枪法测序仍然劳动强度大。这部分是由于在完成的最后阶段需要人工干预。机器视觉技术的进步提高了图谱绘制的通量,并为图谱绘制的完全自动化开辟了道路,从而增强了光学图谱在这个问题上的适用性。对图谱和序列数据进行了比较,这些数据表征了序列间隙并指导了新生组装。