相关实验视频

Updated: Jul 4, 2025

Author Spotlight: Advancements in DNA Nanosensors – Addressing Sensitivity and Selectivity Challenges in Molecular Detection

Author Spotlight: Advancements in DNA Nanosensors – Addressing Sensitivity and Selectivity Challenges in Molecular Detection

Published on: February 9, 2024

探索和应用DNA结合蛋白来制造多功能DNA-蛋白共价链接贴片 (D-Pclip):生物传感元件的案例

Erika Komiya¹, Shouhei Takamatsu¹, Daimei Miura¹

¹Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.

Journal of the American Chemical Society

|January 31, 2024

概括

我们开发了一种DNA-蛋白共价结合贴片 (D-Pclip) 用于稳定,敏感的生物传感. 这种方法可以创建固体测量DNA-蛋白质复合体,在各种条件和样本中改善检测.

科学领域:

生物化学
分子生物学
生物技术

背景情况:

非共价DNA-蛋白质复合体对于生物感知至关重要,但由于环境不稳定,其敏感性较低.
为了提高生物传感器的性能,开发稳定和静态的DNA-蛋白质复合体是必不可少的.

研究的目的:

开发一种用于制造共价和固态测量DNA-蛋白质复合体的多功能方法.
提高基于DNA蛋白质的生物传感元件的稳定性和灵敏性.

主要方法:

使用 uracil-DNA glycosylase (UdgX) 和 SpyTag/ SpyCatcher 系统进行了DNA-蛋白共价结合贴片 (D-Pclip) 的设计.
UdgX因其高DNA结合效率 (> 90%) 和与含有 uracil 的 DNA 形成共价键的能力而被选中.
使用D-Pclip制备了一种血红蛋白 (Hb) 结合的阿胺和葡萄糖氧化酶 (GOx) 的固体测量复合物.

主要成果:

D-Pclip系统在4°C时成功地制造出静态DNA-蛋白质复合体.
使用D-Pclip制备的阿普坦-GOx复合物以剂量依赖的方式检测出Hb.
在缓冲剂和人血清中检测Hb在临床相关范围内是有效的,不需要洗步骤.

结论:

D-Pclip提供了一种通用且方便的方法来制造稳定,静态的DNA-蛋白质复合体.
这项技术在生物传感和其他需要精确的DNA-蛋白质结合的研究领域具有显著的应用潜力.

更多相关视频

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Published on: August 27, 2014

DNA Tension Probes to Map the Transient Piconewton Receptor Forces by Immune Cells

DNA Tension Probes to Map the Transient Piconewton Receptor Forces by Immune Cells

Published on: March 20, 2021

相关概念视频

Labeling DNA Probes

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

DNA Microarrays

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...

Southern Blot

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...