Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.664031
Title: Autonomous maintenance of fission yeast DNA in mouse cells
Author: Wright, Diana
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1993
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Abstract:
This thesis describes an investigation into the structure and function of eukaryotic chromsomes. The experimental approach used was to study the maintenance of Schizosaccharomyces pombe chromosomes introduced into mouse fibroblasts by cell-to-cell fusion. Work on the development of rapid tests for screening hybrid clones and attempts to isolate novel S. pombe / mouse cell hybrids are described in Chapter 3. A detailed study of the structure and location of the fission yeast DNA present in fusion hybrid line F7-2 is presented in Chapters 4 and 5. This hybrid carried a 1.4 Mb fragment of S. pombe chromosome III DNA, confirming previous observations that large pieces of yeast DNA can be introduced into mammalian cells by fusion. Pulsed field gel analysis indicated that this fragment of S. pombe chromosome III DNA was part of a much larger molecule that may have been generated by co-ligation of fragments from several fission yeast chromosomes. S. pombe telomere and telomere-associated sequences were retained in F7-2 indicating that the tips of fission yeast chromosomes can be stabilised in mousecells either by the addition of host cell telomeric repeats or by rearrangements that place them at internal sites in host cell chromatin. A large proportion of the S. pombe DNA in fusion hybrid F7-2 was unstably inherited and is believed to be located on an autonomous extrachromosomal element comparable to the double minute chromosomes observed in drug-resistant mammalian cell lines. Studies on the replication of the S. pombe DNA in hybrid F7-2 are described in Chapter 6. The bulk of the S. pombe DNA was observed to be replicated once per cell cycle indicating that fission yeast DNA contains sites which can act as origins of DBA replicaiton in mouse cells and that initiation of DNA synthesis at these sites is controlled by the host cell mechanism that prevents re-replication of DNA which has already been copied once during S-phase.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.664031  DOI: Not available
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