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Related Experiment Videos

Single M13 bacteriophage tethering and stretching.

Ahmad S Khalil1, Jorge M Ferrer, Ricardo R Brau

  • 1Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

Proceedings of the National Academy of Sciences of the United States of America
|March 16, 2007
PubMed
Summary

M13 bacteriophage displays biomolecules for single-molecule biophysics assays. This viral template

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Area of Science:

  • Biophysics
  • Materials Science
  • Molecular Biology

Background:

  • M13 bacteriophage offers a unique, organized structure for presenting biomolecules.
  • Previous applications focused on directing nanocrystal and materials assembly.
  • This study explores its utility in single-molecule (SM) biophysics.

Purpose of the Study:

  • To develop a novel single-molecule assay utilizing M13 bacteriophage.
  • To quantitatively characterize the elasticity of the M13 biopolymer.
  • To assess the M13 template's potential in tethered bead architectures.

Main Methods:

  • Genetically engineered M13 bacteriophage constructs with hetero-functional ends and capsid display.
  • Development of a single-molecule assay for bead tethering.

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  • Quantitative elasticity characterization using optical trapping with nanometer resolution.

    • Main Results:

    • Successfully demonstrated tethering of microscopic beads using engineered M13 particles.
    • Developed and validated a new single-molecule assay based on M13 bacteriophage.
    • Determined a mean persistence length of ~1,265 nm and a stretching modulus twice that of dsDNA.

    Conclusions:

    • The M13 bacteriophage serves as a versatile platform for single-molecule biophysics assays.
    • Its mechanical properties, including a shorter persistence length than previously thought, are now quantitatively characterized.
    • The M13 template is a promising candidate for advanced tethered bead assays due to its versatility and load-bearing capacity.