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Title: Replication of plasmids containing bovine papillomavirus or Epstein-Barr virus sequences in mammalian cells
Authors: Gahn, Toni A.
Keywords: Molecular biology.
Issue Date: 1990
Publisher: ProQuest Dissertations & Theses
Citation: Source: Dissertation Abstracts International, Volume: 50-11, Section: B, page: 4903.;Advisors: Carl Schildkraut.
Abstract: DNA replication in mammalian cells proceeds with a defined temporal order that depends on cell type. To develop a model to study the timing of replication of mammalian chromosomal DNA, the replication of two viruses, bovine papillomavirus (BPV) and Epstein-Barr virus (EBV) has been examined. BPV transforms rodent fibroblasts while EBV immortalizes human B lymphocytes. In both cases, the viral DNA is maintained predominantly extrachromosomally at a stable copy number.;BPV was found to replicate throughout S phase of the cell cycle. Hybrid plasmids containing the transforming region of BPV plus either the human {dollar}\beta{dollar}-globin or the human interferon {dollar}\beta\sb1{dollar} gene also replicate throughout S. The EBV genome was found to replicate predominantly in the first half of the S phase in an EBV immortalized lymphoblastoid cell line and in one Burkitt's lymphoma line, but either in the middle of or throughout S phase in another Burkitt's lymphoma line. Small plasmids containing the EBV origin of plasmid replication, oriP, were found to replicate either in the middle of or throughout S in both a cell line that contains EBV and in one that does not.;EBV oriP consists of two elements; a region of dyad symmetry and a family of direct repeats. To examine the function of the two elements of oriP, a two dimensional gel electrophoresis technique was employed. An analysis of the branched molecules generated during replication of an oriP plasmid has shown that replication initiates at or very near the dyad symmetry element. The family of direct repeats has been found to contain a barrier to replication and the primary termination site is either within or near the family of repeats. The close proximity of initiation and termination sites in oriP results in replication proceeding in a predominantly unidirectional manner. An analysis of molecules generated by the replication of the EBV genome in an EBV immortalized cell line suggests that the elements of oriP function in a similar manner in the EBV genome.
Appears in Collections:Albert Einstein College of Medicine: Doctoral Dissertations

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