Jove
Visualize
お問い合わせ
JoVE
x logofacebook logolinkedin logoyoutube logo
JoVEについて
概要リーダーシップブログJoVEヘルプセンター
著者向け
出版プロセス編集委員会範囲と方針査読よくある質問投稿
図書館員向け
推薦の声購読アクセスリソース図書館諮問委員会よくある質問
研究
JoVE JournalMethods CollectionsJoVE Encyclopedia of Experimentsアーカイブ
教育
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab Manual教員リソースセンター教員サイト
利用規約
プライバシーポリシー
ポリシー

関連する概念動画

Replication in Eukaryotes02:31

Replication in Eukaryotes

173.0K
Overview
173.0K
The Replisome03:01

The Replisome

35.2K
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...
35.2K
DNA Isolation01:24

DNA Isolation

41.0K
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
41.0K
DNA Replication02:40

DNA Replication

51.2K
DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied.  After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
Replication in Prokaryotes
DNA replication...
51.2K
Replication in Prokaryotes01:32

Replication in Prokaryotes

25.4K
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...
25.4K
Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

15.1K
For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
15.1K

こちらも読む

関連記事

共著者、ジャーナル、引用グラフによってこの研究に関連する記事。

並び替え
Same author

Cryo-EM maps of human DNA polymerase ε should be reevaluated in light of its unexpected behavior in vitro.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

Competition for the nascent leading strand shapes the requirements for PCNA loading in the replisome.

The EMBO journal·2025
Same author

Structural basis for processive daughter-strand synthesis and proofreading by the human leading-strand DNA polymerase Pol ε.

Nature structural & molecular biology·2024
Same author

CST-polymerase α-primase solves a second telomere end-replication problem.

Nature·2024
Same author

CST-Polymeraseα-primase solves a second telomere end-replication problem.

bioRxiv : the preprint server for biology·2023
Same author

Structure of a human replisome shows the organisation and interactions of a DNA replication machine.

The EMBO journal·2023

関連する実験動画

Updated: Sep 22, 2025

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein
07:19

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein

Published on: April 26, 2024

2.8K

精製されたヒトタンパク質で迅速かつ効率的なDNA複製

Yasemin Baris1, Martin R G Taylor1, Valentina Aria1

  • 1MRC Laboratory of Molecular Biology, Cambridge, UK.

Nature
|May 18, 2022
PubMed
まとめ

研究者らはヒトの複製体を再構成し,DNAポリメラーゼエプシロン (Polε) がリードストランド合成の鍵であることを明らかにした. このプロセスはPCNAとCTF18-RFCを必要とし,AND-1はリードストランド複製を直接支援する.

科学分野:

  • 分子生物学
  • 遺伝学
  • 生物化学

背景:

  • ヒトのレプリソームは,DNA複製に不可欠であり,CMGヘリケースの周りに組織された多数のタンパク質で構成されています.
  • 精製されたタンパク質から人間の複素体を完全に再構成することは,その正確な機能の理解を妨げています.

研究 の 目的:

  • 精製した成分を使って 機能的なヒト複製体を再構成する
  • ヒトのDNA複製におけるDNAポリメラーゼと付属因子の特定の役割を解明する.

主な方法:

  • 11の浄化されたヒト複製因子 (43のポリペプチド) を用いたヒト複製体の生化学的復元.
  • DNA複製の速度と効率を測定する検査です
  • PCNA,CTF18-RFC,CLASPIN,TIMELESS-TIPIN,AND-1がレプリソーム機能に果たす役割を調査する.

主要な成果:

  • 迅速かつ効率的なDNA複製を可能にする人間の複製体を再構成しました.
  • PCNAとCTF18-RFCに依存する,最適なリードストランド合成に不可欠なDNAポリメラーゼエプシロン (Polε) を特定した.
  • 酵母と異なり,AND-1はリードストランドの複製を直接強化し,Polαは遅滞ストランドのプリミングのために独立して採用されていることが示されました.

さらに関連する動画

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
08:53

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

Published on: May 2, 2025

497
Author Spotlight: Characterizing DNA Replication of Pathogenic Repeats to Uncover Mechanisms of Replication Fork Stalling and Expansion
05:22

Author Spotlight: Characterizing DNA Replication of Pathogenic Repeats to Uncover Mechanisms of Replication Fork Stalling and Expansion

Published on: September 13, 2024

895

関連する実験動画

Last Updated: Sep 22, 2025

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein
07:19

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein

Published on: April 26, 2024

2.8K
Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
08:53

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

Published on: May 2, 2025

497
Author Spotlight: Characterizing DNA Replication of Pathogenic Repeats to Uncover Mechanisms of Replication Fork Stalling and Expansion
05:22

Author Spotlight: Characterizing DNA Replication of Pathogenic Repeats to Uncover Mechanisms of Replication Fork Stalling and Expansion

Published on: September 13, 2024

895

結論:

  • 再構成されたヒト複製体は DNA複製メカニズムを研究するためのシステムを提供します.
  • ヒトのリードストランド合成におけるPolε,PCNA,CTF18-RFC,AND-1の明確な役割が明らかになった.
  • ヒトのDNA複製における遅滞鎖の原始化のためのPolαの独立した採用を明らかにした.