Long-range and highly sensitive DNase I footprinting by an automated infrared DNA sequencer.
Long-range and highly sensitive DNase I footprinting by an automated infrared DNA sequencer.
复制标题
通过自动红外 DNA 测序仪进行长距离、高灵敏度 DNase I 足迹分析。
DOI：
10.2144/97232st08
复制
发表时间：
1997
期刊：
影响因子：
2.7
通讯作者：
D. Sorensen
中科院分区：
文献类型：
--
作者：
M. Machida;H. Kamio;D. Sorensen
我们已经证明，自动 DNA 测序仪适用于 DNase I 足迹中基于荧光的片段检测。我们展示了利用红外荧光自动 DNA 测序仪进行长距离、高灵敏度 DNase I 足迹分析的潜力。在单次电泳运行中，可重复检测到 895 bp DNA 片段双链上约 100 至 750 bp 的人类转录因子 SpI 足迹。我们开发了用于高灵敏度检测的数据收集和后续图像处理技术。使用 3.1 fmol 的 512 bp DNA 片段检测到小于 0.1 足迹单位（fpu：约 4.5 ng）的 SpI。与之前可见染料荧光 DNA 测序仪报道的灵敏度相比，灵敏度提高了 10 倍以上。该方法对于转录调控区的系统分析和转录过程的大规模分析将非常重要。
We have shown that an automated DNA sequencer is applicable to fluorescence-based detection of fragments in DNase I footprinting. We demonstrated the potential of long-range and highly sensitive DNase I footprinting taking advantage of an infrared-fluorescence automated DNA sequencer. Footprints of human transcription factor SpI were reproducibly detected ranging approximately between 100 and 750 bp on both strands of an 895-bp DNA fragment in a single electrophoresis run. We developed techniques in data collection and subsequent image processing for highly sensitive detection. Less than 0.1 footprinting unit (fpu: approximately 4.5 ng) of SpI was detected using 3.1 fmol of a 512-bp DNA fragment. This is greater than 10-fold increase in sensitivity over what has previously been reported by visible dye fluorescence DNA sequencers. This method will be very important in systematic analysis of transcription regulatory regions and in large-scale analysis of the transcription process.