Methylation analysis of a plasmid containing a mammalian cell cycle regulatory sequence after transient transfection...

A hamster genomic sequence containing a cell cycle regulatory sequence derived from a histone H3.2 gene was transfected into K12 hamster fibroblasts in the form of plasmid DNA prepared from dam+ E. coli. Analysis of the plasmid DNA recovered 72 hr after the transfection revealed that it was resistant to Mbol but was sensitive to Dpn 1 enzyme digestion, suggesting that this plasmid had retained its dam E. coli methylated sites and was therefore unable to undergo replication following transfection into homologous host cells. These results were discussed with relation to a previously described yeast cell cycle regulatory sequence which was functionally linked to an autonomous replicating sequence and was located near a yeast H2B gene.