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指导合成DNA结合分子的设计,使用大规模并行测序
Jordan L Meier1, Abigail S Yu, Ian Korf
1Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Journal of the American Chemical Society
|September 28, 2012
概括
role-imidazole聚胺与高特异性结合DNA. 高通量测序揭示了规范规则预测结合,但也确定了新的地点,使定制的DNA结合分子设计能够用于生物和纳米技术应用.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 化学生物学 化学生物学
背景情况:
- 了解DNA结合分子的特异性对于基因组应用至关重要.
- role-imidazole聚胺是一种设计用于特定序列DNA结合的分子类别.
- 需要高通量方法来绘制大型DNA库中的结合点.
研究的目的:
- 通过使用高通量方法来确定pyrrole-imidazole聚胺的DNA结合特异性.
- 为了确定正规和非正规的高亲和度结合点.
- 为了使新型聚胺具有更好的特异性的合理设计.
主要方法:
- 亲和度净化与大规模并行测序 (深度测序) 相结合.
- 一个10^12个成员的DNA序列库的分析.
- 重新设计的头针聚胺的设计和测试.
主要成果:
- 正规的基配对规则在很大程度上预测了聚胺的DNA结合特异性.
- 在某些聚胺的反向方向中发现了意想不到的高亲和度结合点.
- 重新设计的聚胺成功地区分了类似的DNA序列 (5'-WCGCGW-3'和5'-WGCGCW-3').
结论:
- 高通量测序是一种强大的工具,用于表征DNA结合分子的特异性.
- 罗-意米达聚胺可以合理设计,用于精确的DNA向.
- 这项工作为开发各种应用的序列特定结合分子提供了基础.
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