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相关概念视频

Proofreading01:31

Proofreading

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Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore,  it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...
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Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
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The Replisome03:01

The Replisome

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DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
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Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
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Replication in Eukaryotes02:31

Replication in Eukaryotes

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Overview
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PCR01:32

PCR

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相关实验视频

Updated: Jul 12, 2025

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
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DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

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工程Ca2+-依赖DNA聚合酶活动

Bradley W Biggs1,2, Alexandra M de Paz3,2, Namita J Bhan1,2

  • 1Department of Chemical and Biological Engineering, Northwestern University, Evanston, Illinois 60208, United States.

ACS synthetic biology
|October 19, 2023
PubMed
概括
此摘要是机器生成的。

研究人员设计了DNA聚合酶 (DNAPs) 来检测离子 (Ca2+),克服了基于DNA的生物传感器中的缓慢信号传导. 这一突破使得各种应用的生物传感更快,更响应.

关键词:
生物传感器生物传感器数字化 DNA DNA 数字化 DNA蛋白质工程工程 蛋白质工程

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科学领域:

  • 合成生物学 合成生物学
  • 生物感知技术的技术
  • 分子工程是分子工程.

背景情况:

  • 合成生物学在诊断和遗传编程方面推进了生物传感.
  • 下一代生物传感器需要扩展测量环境,现象和提高质量.
  • 生物传感器中的DNA集成提供了数据记录,但面临信号传输瓶.

研究的目的:

  • 设计DNA聚合酶 (DNAPs) 来感知和响应离子 (Ca2+).
  • 为了克服基于DNA的生物传感器记录中的信号传导限制.
  • 为开发基于DNAP的新型生物传感器奠定基础.

主要方法:

  • 来自Saccharomyces cerevisiae*的Pol δ复制聚合酶复合物的工程组件.
  • 使用域插入和绑定位点移植到Pol δ子单元.
  • 显示对Ca2+的功能性全性敏感性.

主要成果:

  • 成功设计了具有对Ca2+全敏感性的DNAPs.
  • 在经过修改的Pol δ子单位中证明了Ca2+反应性.
  • 建立了一种新的 ligand-gated DNAP 活性方法.

结论:

  • 工程DNAPs可以充当传感器和响应器,对像Ca2+这样的特定连接体起到作用.
  • 这项工作解决了基于DNA的生物传感器中的信号传导瓶问题.
  • 为开发基于DNAP的先进生物传感平台提供了基础.