Methods for measuring the replication and segregation of Epstein-Barr virus-based plasmids.
Plasmids containing the Epstein-Barr virus (EBV) latent origin of replication, OriP, are stably maintained in human cells expressing the viral EBNA-1 protein. This stable maintenance is owing to the ability of EBNA-1 to activate DNA replication from OriP and to facilitate the segregation of the plasmids during cell division. Methods for quantifying the replication and stable maintenance of EBV-based plasmids in human cells are presented here, as is a reconstituted segregation system in yeast that enables the segregation activity of EBNA1 to be measured independently from its replication activity.
Kapoor
Priya
P
Department of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario, Canada.
Frappier
Lori
L
D015139:Blotting, Southern / Q000379:methods*; D002352:Carrier Proteins / Q000235:genetics; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics*; D004854:Herpesvirus 4, Human; D006801:Humans; D010957:Plasmids; D016601:RNA-Binding Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins
D002352:Carrier Proteins; C432687:EBNA1BP2 protein, human; C499008:EBP2 protein, S cerevisiae; D019309:Epstein-Barr Virus Nuclear Antigens; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; C417029:EBV-encoded nuclear antigen 1