The DNA of nuclear polyhedrosis virus (AcNPV) was isolated from extracellular non-occluded virions. The guanosine plus cytosine content of AcNPV DNA was estimated from its buoyant density in CsCl and its T value to be about 43 mol %. In ethidium bromide-CsCl gradients double-stranded covalently closed DNA molecules (form I) were detected, while relaxed circular (form II) and linear (form III) DNA molecules were demonstrated by velocity sedimentation in sucrose. The mol. wt. of AcNPV DNA was calculated to be about 78 × 10 from the sedimentation value of form III DNA. Digestion of AcNPV DNA with restriction endonucleases RI and I resulted in 21 and 7 fragments, respectively, after analysis on agarose gels. The mol. wt. of the fragments and their molar ratio were determined. By this method the mol. wt. of AcNPV DNA was calculated to be about 83 × 10.


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