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関連する概念動画

Translesion DNA Polymerases02:10

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
Proofreading01:31

Proofreading

Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore,  it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase Enzyme
Proofreading01:43

Proofreading

Synthesis of new DNA molecules starts when DNA polymerase links nucleotides together in a sequence that is complementary to the template DNA strand. DNA polymerase has a higher affinity for the correct base to ensure fidelity in DNA replication. The DNA polymerase furthermore proofreads during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.Errors during Replication Are Corrected by the DNA Polymerase EnzymeGenomic DNA is synthesized in...
Lagging Strand Synthesis01:59

Lagging Strand Synthesis

During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...

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関連する実験動画

Updated: Jun 20, 2026

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
07:38

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

Published on: October 6, 2017

ハイドロフォビック塩基の類似体のためのポリメラーゼを進化させる

David Loakes1, José Gallego, Vitor B Pinheiro

  • 1MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, United Kingdom.

Journal of the American Chemical Society
|September 26, 2009
PubMed
まとめ

研究者らは,誘導進化を用いた新しいDNAポリメラーゼ5D4を開発し,水害性塩基類似体 (HBA) を効率的に複製した. このブレークスルーは,新しいアプリケーションのための核酸化学とコーディングの可能性を拡大します.

科学分野:

  • バイオケミストリー バイオケミストリー
  • 分子生物学は分子生物学である.
  • 合成生物学 合成生物学とは

背景:

  • 水嫌塩基類型 (HBAs) は,核酸の化学的およびコーディングの可能性を拡大しています.
  • しかし,HBAは通常,DNAポリメラーゼの劣った基質であり,その応用を妨げています.
  • 適した基板特性を持つHBAを発見することは,大きな課題でした.

研究 の 目的:

  • DNAポリメラーゼの誘導進化によってHBA基板特性を改善するための戦略を開発する.
  • 特定のHBAを複製できるポリメラーゼを選択するために,区画化された自己複製 (CSR) を利用する.
  • 様々なHBAを利用する能力を強化したポリメラーゼを特定する.

主な方法:

  • Thermus属から派生したキメリックDNAポリメラーゼの方向進化.
  • 選択基質として5-ニトロインドール (d5NI) と5-ニトロインドール-3-カルボキシアミド (d5NIC) を使用した区画化された自己複製 (CSR).
  • バイオケミカルアッセイとNMRスペクトロスコピーを用いて,ポリメラーゼ活性,基板特異性,および忠誠性の特徴付け.

主要な成果:

  • 新種のDNAポリメラーゼ5D4を分離し,HBAを利用する能力が大きく向上した.

さらに関連する動画

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization
15:22

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

Published on: April 3, 2014

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
05:37

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

Published on: April 4, 2025

関連する実験動画

Last Updated: Jun 20, 2026

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
07:38

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

Published on: October 6, 2017

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization
15:22

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

Published on: April 3, 2014

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
05:37

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

Published on: April 4, 2025

  • 5D4は,効率的に形成され,拡張されたd5NIとd5NICの自己ペアと,すべての標準ベースを持つヘテロペア.
  • ポリメラーゼ5D4は,さまざまなHBAペアで活性を示し,さまざまなHBAをバイパスし,HBAを含むプライマーのPCR増幅を高精度で可能にしました.

    • 結論:

    • 誘導進化アプローチにより,HBA利用が著しく改善されたポリメラーゼ (5D4) が得られました.
    • 5D4は,複製とDNA合成に適した核塩基アナログのレパートリーを拡張します.
    • このエンジニアリングされたポリメラーゼは,化学的および機能的多様性を拡大した核酸ポリマーを作成するための約束を保持しています.