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Base Excision Repair

Base Excision Repair

One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...

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Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction

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Published on: October 6, 2022

使用外核酶消化加速后SELEX Aptamer工程

Juan Canoura¹, Haixiang Yu¹, Obtin Alkhamis¹

¹Department of Chemistry and Biochemistry, Florida International University, 11200 Southwest Eighth Street, Miami, Florida 33199, United States.

Journal of the American Chemical Society

|December 30, 2020

概括

此摘要是机器生成的。

使用外核酶的新方法可以快速评估胺基结合亲和力,改善胺基工程. 这种技术增强了对像腺这样的特定点的适合体选择,使敏感的生物传感器能够发育.

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科学领域:

生物技术
分子生物学
分析化学

背景情况:

通过指数式丰富的连接体系统演化 (SELEX) 用于分离体,但识别具有高亲和度和特异性的最佳序列是具有挑战性的.
现有的SELEX后的aptamer工程方法往往有局限性,包括偏差,可变的成功率和需要专门的设备.

研究的目的:

开发一种可通用,快速,无标签的测定方法,用于检测阿普坦的结合性质.
设计具有改进的结合特性的体,特别是增强对目标分子的亲和力和特异性.
为了证明开发的方法对小分子和蛋白质结合的体的适用性.

主要方法:

使用外核酶III和外核酶I来通过监测连接体结合时的消化动力学变化来分析体的结合特性.
将试验应用于与外核酶消化动力学相关联的甲基结合DNA胺及其突变物.
通过选突变物和识别高亲和性腺特异性细胞因子,设计了一种与ATP,ADP,AMP和腺无差别结合的DNA细胞因子.

主要成果:

联体结合改变了外核酶消化动力学,这种方式与体-联体亲和关系密切相关.
鉴定出两种高亲和度的体, 特别与突变物库中的腺结合.
开发了一种基于aptamer的电化学传感器,使用工程化aptamers,在50%的血清中达到1μM的检测极限.

结论:

开发的基于外核酶的试验是一种可通用和有效的方法,用于表征胺-联体结合亲缘关系.
这种方法促进了具有改进的结合性质的aptamers的工程,适合各种应用,包括生物传感.
该方法可适应高吞吐量查,可应用于多种序列,结构和长度的DNA体.