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Title: The amplification of the Chinese hamster ovary glutamine synthetase gene and its flanking DNA
Author: Hayward, Bruce
Awarding Body: University of Glasgow
Current Institution: University of Glasgow
Date of Award: 1987
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By multistep selection of cells for increasing resistance to methionine sulphoximine, a specific inhibitor of glutamine synthetase, an amplified cell line was produced with approximately 1000 copies of the glutamine synthetase (GS) gene. The genomic GS gene was isolated from the DNA of this cell line and the cloned DNA used to probe the regions flanking the GS gene. By conventional agarose gel electrophoresis it was shown that the DNA containing the GS gene was homogeneous for at least 40 kbp in all copies of the GS amplification unit and that the hypomethylation patterns at the 5' of the GS gene were also homogeneous. An estimate of the size of the amplified unit suggested that it was 60 kbp in size. Pulse field gel electrophoresis was used to analysis the relationship of one unit to another and several possible models are discussed. PFGE suggested that a single unit was 75 kbp in size and analysis of other related cell lines suggested that this size is a reduction from the size of the unit in a less amplified line. Karyotypic and in situ hybridization studies of the parental and highly amplified cell line revealed the presence of several rearranged marker chromosomes one of which was shown to be the site of the amplified DNA.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available