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Isolation of Herpes Simplex Virus DNA from Virus Particles and Infected Cells by Electrophoresis in Polyacrylamide Gels

Laboratory for Molecular Virology, Hebrew University-Hadassah Medical School, Jerusalem, Israel

The DNA molecules synthesized in herpes simplex virus (HSV) infected cells were analysed by electrophoresis in polyacrylamide gels. It was found that part of the virus DNA, isolated from infected nuclei, migrated into the gels in a manner similar to the virus DNA molecules isolated from purified herpes simplex virus particles. Most of the virus DNA molecules, labelled in vivo for 60 min with [³H]-thymidine as well as labelled cellular DNA molecules, were retained at the top of the gel during electrophoresis. The amount of HSV-DNA which migrated into the gel gradually increased when the labelled cells were reincubated after removal of the isotope (chase). Most of the DNA molecules, which were retained at the top of the gel, had the density of virus DNA but differed in their sedimentation behaviour in sucrose gradients from the intact DNA molecules isolated from purified virus particles. The presence of replicating as well as mature virus DNA molecules in infected nuclei is discussed.

Received 13 March 1974; accepted 6 June 1974.