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Replication in Prokaryotes02:35

Replication in Prokaryotes

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Overview
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Replication in Prokaryotes01:32

Replication in Prokaryotes

28.4K
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.
Many Proteins Work Together to Replicate the Chromosome
Replication is coordinated and carried out by a host of specialized...
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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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Mutations in Microorganisms01:18

Mutations in Microorganisms

868
Mutations are heritable changes in an organism’s genome involving alterations in the base sequence of DNA or RNA. These changes can influence cellular processes and phenotypic traits, potentially transforming the unaltered wild type into a mutant form. Such changes, termed forward mutations, are pivotal in shaping the genetic diversity of organisms.RNA viruses exhibit the highest mutation rates due to the absence of robust proofreading mechanisms during genome replication. In contrast,...
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Mismatch Repair01:20

Mismatch Repair

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Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
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DNA Bacteriophages01:26

DNA Bacteriophages

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Bacteriophages, or phages, are viruses that specifically infect bacteria, utilizing their genetic material to hijack host cellular machinery for replication. DNA bacteriophages employ single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) genomes. These phages exhibit diverse replication strategies and host interactions, influencing their ecological roles and applications in biotechnology and medicine.ssDNA BacteriophagesssDNA phages, with their small genomes, utilize unique strategies to...
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Updated: Feb 26, 2026

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

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大腸菌におけるΦ29ベースの直交複製システムを用いた高変異原性連続進化

Fabian B H Rehm1, Kim C Liu2, Rongzhen Tian2

  • 1Medical Research Council Laboratory of Molecular Biology, Cambridge, UK. frehm@mrc-lmb.cam.ac.uk.

Nature biotechnology
|February 24, 2026
PubMed
まとめ
この要約は機械生成です。

バクテリオファージΦ29成分を用いた安定したDNA複製システムを大腸菌で構築し、遺伝子進化を加速させた。このシステムは効率的に突然変異を導入し、新しい遺伝子機能の急速な発達を可能にする。

キーワード:
直交DNA複製遺伝子進化大腸菌バクテリオファージΦ29高変異原性タンパク質工学合成生物学

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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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09:01

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Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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科学分野:

  • 分子生物学
  • 合成生物学
  • 遺伝学

背景:

  • 加速遺伝子進化には、オフターゲット効果のない精密な超変異が必要である。
  • 既存の遺伝子進化システムは、効率と安定性に限界がある。

研究 の 目的:

  • 大腸菌における加速遺伝子進化のための直交DNA複製システムの開発と最適化。
  • 標的遺伝子改変のための高変異原性DNAポリメラーゼの工学的設計。

主な方法:

  • バクテリオファージΦ29の構成要素を利用して、最小限の直交DNA複製システムを作成した。
  • in vivoで複製子を工学的に設計し、高変異原性のΦ29 DNAポリメラーゼを開発した。
  • 数百世代にわたってシステムの安定性を維持した。

主要な成果:

  • 10^-4/塩基/世代に迫る突然変異頻度を達成した。
  • チゲサイクリンに対するテトラサイクリン耐性の急速な進化を実証した。
  • 3日間で第三世代セファロスポリンに対するβ-ラクタマーゼ活性を1,000倍に増加させた。

結論:

  • 開発されたΦ29ベースのシステムは、遺伝子機能の安定した、連続的で、加速された進化を可能にする。
  • このシステムは、新しいまたは改善された遺伝子特性の工学的設計の速度と有効性を大幅に向上させる。
  • 合成生物学およびタンパク質工学の応用にとって強力なツールを提供する。