Conductance Fluctuations: A New Approach to Sequencing?

项目摘要 这个为期两年的R21项目的目标是探索一种非常新的方法来探索 基于最近发现的蛋白质的电子性质的酶活性。长期目标是创造 一种低成本、高速的单分子基因组测序仪，具有长读取，不需要染料或标记，具有 直接电子读数。这项技术具有快速阅读的潜力(大约每小时一小时 基因组)使用集成电路芯片。如果要充分实现这一潜力，那么利用基因组 在临床上进行近乎实时反馈的测序可能成为现实。初始数据显示高信号 对蛋白质波动的噪声测量似乎是可能的，并表明至少有一些 波动与蛋白质的功能有关。然而，这些研究受到很大的限制。 由于与蛋白质接触的化学性质的不确定性，导致了可变性的增加。因此，我们的首要目标是 是设计具有内置接触的聚合酶，设计成自组装成测量设备。我们 将用扫描隧道显微镜测试该结构，并在此过程中获得设计参数 一种测序装置。我们的第二个目标是测试固态设备中的至少一个结构，以检查 扫描隧道显微镜中获得的数据的有效性。如果成功，收集的数据将为 为扩大规模和提高准确性以实现可行的测序平台奠定基础 上述属性。

Project Summary The objective of this two-year R21 project is to explore a very new approach to probing the dynamics of enzyme activity based on recently-discovered electronic properties of proteins. The long-term goal is to create a low-cost, high speed single-molecule genome sequencer with long reads, requiring no dyes or labels, with direct electronic readout. The technology has the potential for rapid reads (on the order of an hour per genome) using an integrated circuit chip. If this potential were to be fully realized, then use of genome sequencing in the clinic with near real-time feedback could become a reality. Initial data shows that high signal to noise measurement of protein fluctuations appears to be possible, and suggests that at least some of the fluctuations are associated with the function of the protein. However, these studies are limited by a great deal of variability owing to uncertainty about the chemical nature of contacts to the proteins. Therefore, our first aim is to engineer polymerases with built-in contacts, designed to self-assemble into a measurement device. We will test the construct with a scanning tunneling microscope, and, in the process, obtain design parameters for a sequencing device. Our second aim is to test at least one structure in a solid-state device, to check the validity of data obtained in a scanning tunneling microscopy. If successful, the data collected would lay the groundwork for a larger effort to scale up and improve accuracy to achieve a viable sequencing platform with the properties described above.