The Characterization of an in Vitro Isolated Nuclei System for the Investigation of the Mechanism of Herpesvirus DNA Replication in Infected Human Embryonic Lung Cells
Infection of human embryonic lung (HEL) cells by Herpes simplex type II virus stimulates labeled thymidine incorporation into an acid-preeipitable product as much as twentyfold higher than by mock-infection. The rates of overall in vitro and in vitro DNA synthesis and the relative fractions of Herpes and cell DNA synthesized in vitro and in vitro by nuclei isolated from the infected cells are the same at various times after infection. All four rNTPs strikingly stimulate 3H-TTP incorporation in nuclei from Herpes-infected but not mock-infected cells. The thermal lability of in vitro DNA synthesis is different for Herpes-infected than for mock-infected cells, although the relative fractions of cell and viral DNA made at 42°C and at 34°C are the same. Results of variable time pulse label experiments with the isolated nuclei system suggest a discontinuous mode of DNA replication.
KeywordsHerpes Simplex Type Human Embryonic Lung Isotonic Buffer Human Embryonic Lung Cell Roux Flask
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