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相关概念视频

Replication in Prokaryotes01:32

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DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
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In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
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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.
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Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
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RoCi - 一个基于滚动循环复制的单步多副本集成系统.

Martzel Antsotegi-Uskola1, Vasil D'Ambrosio2, Brian King Himm Mo2

  • 1Department of Biotechnology and Biomedicine, Technical University of Denmark, Section for Synthetic Biology, 2800 Kongens Lyngby, Denmark.

Nucleic acids research
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PubMed
概括
此摘要是机器生成的。

我们开发了RoCi,这是一种将真菌中的多个基因拷贝集成的新方法,增强了酶和代谢物生产. 这种滚动循环复制技术简化了用于工业应用的细胞工厂建设.

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科学领域:

  • 分子生物学分子生物学
  • 合成生物学 合成生物学
  • 生物技术是生物技术.

背景情况:

  • 菌是生产酶和代谢物的关键细胞工厂.
  • 产生高产的真菌菌株往往需要整合多个基因拷贝,这是一个挑战.
  • 目前用于多个基因整合的现有方法通常是复杂和低效的.

研究的目的:

  • 开发一种简单有效的方法,将多个基因表达录音带集成到真菌中定义的基因组位点中.
  • 改进真菌细胞工厂的建设,以提高代谢物和酶的生产.
  • 为了证明该方法在不同真菌物种中的适用性.

主要方法:

  • 开发了RoCi (滚动圆集成),一种利用滚动圆复制进行基因盒集成的方法.
  • 在不需要基于大肠杆菌的克隆的情况下执行RoCi,从而实现自动化实验.
  • 在Aspergillus nidulans中应用RoCi,以产生多达70个mRFP副本的菌株.

主要成果:

  • RoCi促进了多拷贝基因阵列的稳定集成,支持在固体介质中至少150代高稳定的mRFP生产.
  • 通过将基因表达盒优化为有缺陷的促进器控制选择标记,在液体培养中减轻了轻微的拷贝损失.
  • 与单复制品菌株相比,RoCi的应用导致cordicepin和β-glucuronidase的产量增加了约10倍.
  • 在工业真菌劳动马中成功应用RoCi 黑色菌和小菌.

结论:

  • RoCi是一种简单,高效和多用途的方法,用于在真菌中集成多个基因副本.
  • 该方法显著提高了真菌细胞工厂生产有价值的代谢物和酶的产量.
  • 对于合成生物学和工业生物技术的应用,RoCi具有前景,包括A. niger和A. oryzae.