Measurements of DNA lengths remaining in a viral capsid after osmotically suppressed partial ejection.
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.
Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095-1569, USA.
Alex
A
Evilevitch
James W
JW
Gober
Martin
M
Phillips
Charles M
CM
Knobler
William M
WM
Gelbart
D000042:Absorption; D000956:Antigens, Viral / Q000737:chemistry*; D010582:Bacteriophage lambda / Q000378:metabolism; D002213:Capsid / Q000737:chemistry*; D020285:Cryoelectron Microscopy; D004247:DNA / Q000737:chemistry*; D003850:Deoxyribonuclease I / Q000737:chemistry; D004586:Electrophoresis; D016521:Electrophoresis, Gel, Pulsed-Field; D004926:Escherichia coli / Q000378:metabolism; D009995:Osmosis; D009997:Osmotic Pressure; D019800:Phenol / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D014466:Ultraviolet Rays
D000956:Antigens, Viral; D019800:Phenol; D011092:Polyethylene Glycols; D004247:DNA; D003850:Deoxyribonuclease I