SV40 DNA EXTRACTED FROM PERSISTENTLY INFECTED VERO CELLS USING MINIPREP COLUMNS FOR PLASMIDS

A recombinant pAT153 plasmid carrying the whole simian virus 40 (SV40) genome in the form of two consecutive Pst I restriction fragments was rapidly isolated using a commercially available minipreps DNA purific ation system. This rapid and simple method was used to extract SV40 DN A from infected Vero monkey cells. Viral DNA replication in persistent ly infected SVP-1 monkey cells was also conveniently followed over a p eriod of 8 d by agarose gel electrophoresis and molecular hybridizatio n using minipreps and the recombinant plasmid (pLCB104) as a probe. Un satisfactory results were obtained, however, when herpes simplex virus DNA was tentatively extracted from infected Vero cells with the above method. Only covalently closed circular DNA molecules, with two stran ds unable to separate fully under denaturing conditions, were apparent ly retained after rapid neutralization by the silicone-based minipreps DNA purification resin.