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関連する概念動画

Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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DNA Agarose Gel Electrophoresis02:35

DNA Agarose Gel Electrophoresis

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Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
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Restriction Enzymes01:11

Restriction Enzymes

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Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
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Fixing Double-strand Breaks02:04

Fixing Double-strand Breaks

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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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Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

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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...
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Southern Blot02:57

Southern Blot

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Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
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Updated: Oct 9, 2025

Author Spotlight: Characterizing Novel Enzymes from Extremophiles and Common Pathogens to Understand DNA Repair and Replication
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Published on: July 5, 2024

876

DNAとDNAを結合する.

Alavattam Sreedhara1, Yingfu Li, Ronald R Breaker

  • 1Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520-8103, USA.

Journal of the American Chemical Society
|March 18, 2004
PubMed
まとめ
この要約は機械生成です。

研究者らは,触媒DNAを用いた新しいDNA結合システムを開発し,T4DNAリガゼのようなタンパク質酵素の必要性を排除した. このデオキシリボ酵素リガゼシステムは,酵素プロセスを模倣して,効率的なDNA結合を達成します.

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In vitro Assembly of Semi-artificial Molecular Machine and its Use for Detection of DNA Damage
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In vitro Assembly of Semi-artificial Molecular Machine and its Use for Detection of DNA Damage

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Associated Chromosome Trap for Identifying Long-range DNA Interactions
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Associated Chromosome Trap for Identifying Long-range DNA Interactions

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関連する実験動画

Last Updated: Oct 9, 2025

Author Spotlight: Characterizing Novel Enzymes from Extremophiles and Common Pathogens to Understand DNA Repair and Replication
05:33

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Published on: July 5, 2024

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In vitro Assembly of Semi-artificial Molecular Machine and its Use for Detection of DNA Damage
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In vitro Assembly of Semi-artificial Molecular Machine and its Use for Detection of DNA Damage

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科学分野:

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

背景:

  • DNA結合は,分子クローニングに不可欠であり,通常はT4DNAリガゼのようなタンパク質酵素に依存します.
  • T4DNAリガゼは,DNA断片の結合における重要なステップである,フォスフォディエステル結合形成を触媒化するためにATPを必要とします.

研究 の 目的:

  • タンパク質酵素なしでDNA結合を触媒にできるDNA配列を分離し,特徴づけること.
  • 触媒DNA (デオキシリボジーム) を用いた新しい,酵素のないDNA結合システムを開発する.

主な方法:

  • リガゼ活性を持つDNA配列を特定するために,インビトロ選択が採用されました.
  • ATPに依存する自己アデニル化デオキシリボ酵素 (AppDNA) と自己結合性デオキシリボ酵素を含む2段階のプロセスが利用されました.
  • 結合ステップの前に,中間物質の浄化が行われました.

主要な成果:

  • タンパク質酵素がない場合でもDNA結合を触媒化するDNA配列を分離した.
  • デオキシリボ酵素の結合系は,3',5'-フォスフォディエステル結合を形成し,T4DNAリガゼを模倣する.
  • 最適化されたデオキシリボ酵素リガゼは,初期速度定数 (k(obs)) 1 x 10(-4) min(-1 を示し,単純なDNAテンプレートよりも少なくとも10(5) 倍速くリガーション速度を達成しました.
  • 分離されたオリゴヌクレオチドを結合するためのトランス作用構造が開発されましたが,配列の制限があります.

結論:

  • 触媒DNAはDNA断片を効果的に結合させ,タンパク質ベースの酵素の代替品を提供することができます.
  • この酵素なしの結合系は,合成生物学や分子クローンにおける応用の可能性を秘めている.
  • より広範な適用性のために,シーケンス固有の制限を克服するために,さらなる最適化が必要になる可能性があります.