By A. J. MacGillivray
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Electron microscopic examination is essential. There is no agreed marker enzyme for the nuclear envelope, a high specific activity of which implies a pure and intact preparation. The intrinsic error in measuring the nuclear-envelope-specific poly(G)-stimulated nucleoside triphosphatase (Agutter, Harris and Stevenson, 1977) is too great for marker purposes and recent evidence (Comerford et al, 1985) indicates that the 'nuclear envelope peroxidase' (Stubbs and Harris, 1978; Milne et al, 1978) is probably an intranuclear enzyme in vivo that becomes associated with nuclear envelope preparations by spontaneous thiol group oxidation in the presence of traces of Ca (cf.
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8) or polyadenylating activities (D. A. Jackson, unpublished). Nucleoid-associated phosphorylation and methylation activities can be demonstrated using [γ-32Ρ]ΑΤΡ and S-adenosyl-L-[methyl-3H]methionine (SAM) as the respective phosphate and methyl donors. 7. 0 and ImM EDTA; this second centrifugation is used routinely to give a high concentration of nucleoids in a low salt buffer. Under these conditions labelling is linear over a period of 30 min and is approximately 25% of the rate found in an equivalent number of nuclei.