Wu Tian, Schwartz David C
Laboratory for Molecular and Computational Genomics, Department of Chemistry, Laboratory of Genetics, University of Wisconsin-Madison, Madison, WI 53706, USA.
Anal Biochem. 2007 Feb 1;361(1):31-46. doi: 10.1016/j.ab.2006.10.042. Epub 2006 Nov 16.
A new single-molecule system, Transchip, was developed for analysis of transcription products at their genomic origins. The bacteriophage T7 RNA polymerase and its promoters were used in a model system, and resultant RNAs were imaged and detected at their positions along single template DNA molecules. The Transchip system has drawn from critical aspects of Optical Mapping, a single-molecule system that enables the construction of high-resolution ordered restriction maps of whole genomes from single DNA molecules. Through statistical analysis of hundreds of single-molecule template/transcript complexes, Transchip enables analysis of the locations and strength of promoters, the direction and processivity of transcription reactions, and the termination of transcription. These novel results suggest that the new system may serve as a high-throughput platform to investigate transcriptional events on a large genome-wide scale.
一种名为Transchip的新型单分子系统被开发出来，用于在转录产物的基因组起源处对其进行分析。在一个模型系统中使用了噬菌体T7 RNA聚合酶及其启动子，并沿着单模板DNA分子在其位置对产生的RNA进行成像和检测。Transchip系统借鉴了光学图谱的关键方面，光学图谱是一种单分子系统，能够从单个DNA分子构建全基因组的高分辨率有序限制图谱。通过对数百个单分子模板/转录复合物的统计分析，Transchip能够分析启动子的位置和强度、转录反应的方向和持续合成能力以及转录的终止。这些新结果表明，该新系统可能作为一个高通量平台，用于在全基因组范围内大规模研究转录事件。
