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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...
Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
Gene Conversion02:08

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...

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Updated: Jul 6, 2026

Electroeluting DNA Fragments
06:13

Electroeluting DNA Fragments

Published on: September 5, 2010

チメリック金属ペプチドによる配列選択DNA分裂.

Roger T Kovacic1, Joel T Welch, Sonya J Franklin

  • 1Department of Chemistry, University of Iowa, Iowa City, IA 52242, USA.

Journal of the American Chemical Society
|May 29, 2003
PubMed
まとめ
この要約は機械生成です。

この研究では,DNA分裂のために設計された新種の人工核酶,キメリック金属ペプチド (LnP3W) が導入されました. このペプチドは配列選択的なDNA切断を示し,人工酵素設計における重要な進歩を示しています.

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Last Updated: Jul 6, 2026

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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

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科学分野:

  • バイオ・オーガニック化学 バイオ・オーガニック化学
  • ペプチドデザイン ペプチドデザイン
  • 核酸化学 核酸化学について

背景:

  • 人工ヌクレアスの開発は,分子生物学とバイオテクノロジーにとって極めて重要です.
  • キメリックペプチドは,新しい機能的なバイオ分子を設計するためのプラットフォームを提供します.
  • ランタニド金属結合ペプチドは,特定の触媒活動のために設計することができます.

研究 の 目的:

  • 人工ヌクレアゼとして新しいキメリック金属ペプチド (P3W) を設計し,特徴づけること.
  • ランタニド-P3W複合体のDNA結合と分裂活動を調査する.
  • 人工ヌクレアゼによるDNA分裂の配列選択性とメカニズムを評価する.

主な方法:

  • DNA結合モチーフを組み込んだ33メルペプチド (P3W) のペプチド合成と特徴付け.
  • ランタニド (Eu(III),Ce(IV)) とカルシウム (Ca) を用いた金属イオン結合研究.
  • 金属結合時にペプチドの二次構造を決定するための円形二重化スペクトロスコーピー.
  • 超巻きおよび線形化されたプラズミドDNA,およびラベル付けされたDNA断片を用いたDNA分裂アッセイ.
  • DNA分裂産物の分析により,端末とメカニズムを決定する.

主要な成果:

  • P3Wペプチドはランタニドとカルシウムと結合し,EFハンドペプチドに特有の条件形成定数を示します.
  • 金属結合のP3W (EuP3W,CeP3W) は,アルファヘリクスの含有量が大きく,金属結合の成功折り合いを示しています.
  • EuP3WとCeP3Wは,スーパーコイルされたDNAを効果的にニックし,線形化されたDNAを割りますが,EuP3WはpH 8.0でより活発です.
  • CeP3WによるDNA割れは,独占的に3'-OPO(3) と5'-OPO(3) 末端を生成し,地域選択的メカニズムを示唆しています.
  • メタロペプチドは,自由金属イオンとは異なり,DNA分裂において控えめな配列差別を示す.

結論:

  • 新しく設計されたLnP3W金属ペプチドは,活性で配列選択的な人工ヌクレアゼとして機能します.
  • これは,核酵素活性と配列特異性を組み合わせた最初の小さな,非派生性ペプチドを表しています.
  • 折りたたまれた金属ペプチドメインは,おそらくDNA結合と選択的な分裂を媒介する.