Use of Light to Footprint DNA in Vitro and in Vivo

这项研究的最终目标是在完整的细胞中可视化任何基因的结构。当基因表达在转录水平上受到调控时，基因结构的变化会引起基因表达的变化。基因结构上的这些功能差异是由调控蛋白和染色体蛋白与基因的相互作用引发和维持的，也可能是由DNA螺旋结构改变的形成引起的。贝克尔博士正在开发一套光化学方法，这些方法结合在一起，可以让人们检测并指定基因结构中负责调节其活动的那些元素。这种方法是基于早期的工作，该工作证明紫外线可以在单核苷酸分辨率下检测整个细胞中的蛋白质DNA接触。通过在体内而不是在体外直接探测基因结构，可以避免关于基因结构在调节基因表达中的作用的可能不准确或不完整的结论。快速测序任何基因的能力已经彻底改变了生物学。在这里，下一个层次的组织，基因的结构，受到了攻击。通过确定蛋白质- DNA相互作用和基因内改变的DNA结构，将揭示使遗传信息具有生物活性、稳定性和可遗传性的一般组织原理。

The ultimate goal of this research is to visualize, in intact cells, the structure of any gene. When gene expression is regulated at the level of transcription, changes in the structure of the gene are responsible for inducing changes in gene expression. These functional differences in gene structure are initiated and maintained by the interaction of regulatory proteins and chromosomal proteins with the gene, and possibly, by the formation of altered structures within the DNA helix. Dr. Becker is developing a set of photochemical methods, which, when taken together, is allowing one to detect as well as specify those elements of a gene's structure that are responsible for regulating its activity. This approach is based on earlier work which demonstrated that ultraviolet light can detect, at single nucleotide resolution, protein DNA contacts in whole cells. By directly probing gene structure in vivo rather than in vitro, potentially inaccurate or incomplete conclusions regarding the role of gene structure in regulating gene expression will be avoided. The ability to rapidly sequence any gene has revolutionized biology. Here the next level of organization, the structure of the gene, is being attacked. By determining the protein - DNA interactions and altered DNA structures within a gene general organizational principles for rendering the genetic message biologically active, stable, and inheritable will be uncovered.