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相关概念视频

Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

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In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
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Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
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通过DNA定向固定将酶附着在玻璃表面的选择性氧化合反应

Kanwal S Palla1, Tyler J Hurlburt1,2, Alexander M Buyanin1,3

  • 1Department of Chemistry, University of California , Berkeley, California 94720-1460, United States.

Journal of the American Chemical Society
|December 22, 2016
PubMed
概括

我们开发了一种以DNA为导向的固定平台, 这种方法允许酶保持催化活性,并使可重复使用的表面用于生物催化.

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科学领域:

  • 生物化学
  • 材料科学
  • 分子生物学

背景情况:

  • 酶具有很高的选择性和催化率.
  • 固定酶 (异质化) 便于它们的研究,使用和整合到受控系统中.

研究的目的:

  • 开发一个简单的平台,使用DNA定向的蛋白质固定.
  • 验证固定表面的酶活性和可重复使用性.

主要方法:

  • 用单链DNA进行表面修饰.
  • 通过氧化合将补充DNA附加到蛋白质N端.
  • 在DNA修饰的表面上混合蛋白-DNA结合物.
  • 使用光报道器和原子力显微镜进行表征.
  • 在固定阿尔多酶上进行酶活性测定.

主要成果:

  • 在DNA修饰的玻璃表面上成功地定向了酶.
  • 通过固定酶证明了催化活性的保留.
  • 在多个周期中确认了固定表面的可重复使用性.

结论:

  • 通过DNA定向固定提供了一个简单有效的定向蛋白附着方法.
  • 开发的平台保留了酶功能,并允许可重复使用的生物催化表面.