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Title: S. pombe artificial chromosome (SPARC) vectors for cloning large DNA fragments
Author: Young, Dorothy J. D.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1997
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The fusion yeast, Schizosaccharomyces pombe, has several potential advantages as a host for a cloning system for large DNA molecules when compared to the budding yeast, Saccharomyces cerevisiae. The possibility of constructing a large DNA cloning system in S. pombe has been investigated. S. pombe has 3 large chromosomes (greater than 3.5 Mb) and may therefore have the ability to carry megabase sized artificial chromosomes. In addition artificial chromosomes of less than 3.5 Mb will migrate below the host chromosomes on pulsed field gels and thus will be easier to separate away from the host chromosomes. As in higher eukaryotes, the S. pombe centromeres have arrays of repetitive elements, therefore such sequences from heterologous sources which rearrange in S. cerevisiae may not do so in S. pombe. It has been shown that small acentric linearised plasmids with cloned S. pombe telomeres at each end replicate intact in S. pombe. These plasmids were used to prepare vector arms each having a selectable marker, S. pombe telomere and S. pombe replication origin. The S. pombe artificial chromosome (SPARC) vectors do not contain any S. pombe centromere sequences as they are not necessary for the maintenance of the SPARCs under selection. To test the potential of the SPARC vectors for cloning human sequences in S. pombe, well characterised human fragments derived from cosmids were cloned in S. pombe using these SPARC vectors.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available