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Related Concept Videos

The Replisome03:01

The Replisome

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 the...
The Replisome03:01

The Replisome

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 the...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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...
DNA Bacteriophages01:26

DNA Bacteriophages

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...
Homologous Recombination02:31

Homologous Recombination

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...
Coordination of Gene Expression Processes in Bacteria01:29

Coordination of Gene Expression Processes in Bacteria

The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...

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Related Experiment Video

Updated: Jul 17, 2026

Hybrid Ensemble and Single-molecule Assay to Image the Motion of Fully Reconstituted CMG
10:11

Hybrid Ensemble and Single-molecule Assay to Image the Motion of Fully Reconstituted CMG

Published on: July 26, 2024

Single-molecule studies of complex systems: the replisome.

Antoine M van Oijen1

  • 1Harvard Medical School, Dept. of Biological Chemistry and Molecular Pharmacology, 240 Longwood Avenue, Boston, MA 02115, USA. antoine_van_oijen@hms.harvard.edu

Molecular Biosystems
|January 25, 2007
PubMed
Summary

Single-molecule techniques offer unprecedented insights into enzyme dynamics, moving beyond traditional methods. This approach enables the study of complex biological systems like DNA replication, revealing intricate multi-protein interactions.

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Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase
07:37

Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase

Published on: September 27, 2024

Related Experiment Videos

Last Updated: Jul 17, 2026

Hybrid Ensemble and Single-molecule Assay to Image the Motion of Fully Reconstituted CMG
10:11

Hybrid Ensemble and Single-molecule Assay to Image the Motion of Fully Reconstituted CMG

Published on: July 26, 2024

Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase
07:37

Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase

Published on: September 27, 2024

Area of Science:

  • Biochemistry and Molecular Biology
  • Biophysics
  • Systems Biology

Background:

  • Traditional biochemical and molecular biology techniques provide ensemble-averaged data, limiting understanding of dynamic molecular behavior.
  • Quantitative understanding of life's processes relies on kinetics and thermodynamics of biochemical reactions.
  • Ensemble averaging in conventional methods obscures the dynamic behavior of individual molecular components.

Purpose of the Study:

  • To review advances in single-molecule techniques for studying biological processes.
  • To highlight the contribution of single-molecule methods to understanding enzyme dynamics and reaction mechanisms.
  • To illustrate the application of single-molecule techniques to complex multi-protein systems using DNA replication as an example.

Main Methods:

  • Development and application of advanced imaging and molecular manipulation techniques.
  • Observation of individual enzyme activity and recording of 'molecular movies'.
  • Utilizing single-molecule approaches to analyze the orchestration of enzymatic processes in large biomolecular systems.

Main Results:

  • Single-molecule techniques provide dynamic insights into enzyme mechanisms previously unattainable.
  • Demonstration of the feasibility of studying complex systems like the DNA replication machinery at the single-molecule level.
  • Advancement in understanding the coordinated action of multiple enzymes within larger biological assemblies.

Conclusions:

  • Single-molecule techniques are crucial for a complete, system-level understanding of biological processes.
  • Future research should focus on extending single-molecule applicability to larger, more complex multi-protein systems.
  • The DNA replication machinery serves as a key model for showcasing the power of single-molecule approaches in systems biology.