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相关概念视频

DNA Helicases00:55

DNA Helicases

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DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...
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DNA Topoisomerases02:02

DNA Topoisomerases

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Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
Types and Mechanism of action
Topoisomerases are divided into two main types. ...
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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
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相关实验视频

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Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation
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螺旋体活动调制与按需基于光的规格控制

Dmitriy Bobrovnikov1, Monika A Makurath2,3, Clara H Wolfe1

  • 1Department of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland 21205, United States.

Journal of the American Chemical Society
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PubMed
概括

研究人员设计了具有光控制活动的新型DNA螺旋酶. 这些"光螺旋酶"使用光来切换非活跃和活跃状态,为未来的应用提供精确的DNA解控制.

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科学领域:

  • 生物化学
  • 分子生物学
  • 蛋白质工程

背景情况:

  • 蛋白质工程需要理解结构功能关系.
  • 在DNA代谢中,DNA酶是关键的酶.
  • 通过外部刺激控制蛋白质活性是一个关键挑战.

研究的目的:

  • 设计具有光诱导活性的新型DNA螺旋酶.
  • 实现对DNA解的时间空间控制.
  • 探索光遗传学在酶调节中的潜力.

主要方法:

  • 对DNA酶调节子域的结构分析.
  • 新的酶变体的蛋白质工程.
  • 加入基于亚博的交叉连接剂以进行光诱导的异构化.
  • 使用散装生物化学试验和单分子光学针进行表征.

主要成果:

  • 工程螺旋酶表现出依赖光的形状变化.
  • 亚交联异构化将酶活性切换到非活性和活性状态之间.
  • 展示了使用光线对DNA解的需求控制.
  • 单分子分析证实了光调节的酶功能.

结论:

  • 开发出可切换光的DNA螺旋酶 ("光螺旋酶").
  • 在DNA杂交状态上实现了精确的时空控制.
  • 在需要动态控制核酸相互作用的应用中,光螺旋酶具有前景.