Measurements of DNA lengths remaining in a viral capsid after osmotically suppressed partial ejection

Biophys J. 2005 Jan;88(1):751-6. doi: 10.1529/biophysj.104.045088. Epub 2004 Oct 15.

Abstract

The effect of external osmotic pressure on the extent of DNA ejection from bacteriophage-lambda was recently investigated (Evilevitch et al., 2003). The total length of DNA ejected was measured via the 260-nm absorption by free nucleotides, after opening of the capsids in the presence of varying amounts of polyethylene glycol 8000 and DNase I. As a function of osmolyte concentration, this absorption was shown to decrease progressively, ultimately vanishing completely for a sufficiently high external osmotic pressure. In this work we report the results of both sedimentation and gel analysis of the length of DNA remaining inside the capsids, as a function of osmolyte concentration. It is confirmed in this way that the progressive inhibition of DNA ejection corresponds to partial ejection from all of the capsids.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Absorption
  • Antigens, Viral / chemistry*
  • Bacteriophage lambda / metabolism
  • Capsid / chemistry*
  • Cryoelectron Microscopy
  • DNA* / chemistry*
  • Deoxyribonuclease I / chemistry
  • Electrophoresis
  • Electrophoresis, Gel, Pulsed-Field
  • Escherichia coli / metabolism
  • Osmosis
  • Osmotic Pressure
  • Phenol / chemistry
  • Polyethylene Glycols / chemistry
  • Ultraviolet Rays

Substances

  • Antigens, Viral
  • Phenol
  • Polyethylene Glycols
  • DNA
  • Deoxyribonuclease I