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Title: GC rich islands on the human Y chromosome : an approach to cloning Y-linked genes
Author: Shortle, Vivienne Patricia
Awarding Body: University of London
Current Institution: University College London (University of London)
Date of Award: 1990
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Genetic analysis of the human Y chromosome has been slow despite its central role in sex determination. At present genetic analysis has identified only about ten genes on the Y chromosome, and only three genes (one coding for a cell surface antigen, a second coding for a regulatory zinc finger protein and a third coding for a testis specific transcript) have been cloned. This study has attempted to identify further genes on the Y chromosome using techniques which also facilitate cloning. This thesis therefore describes an investigation into the GC rich regions (HTF - Hpall Tiny Fragment Islands) of the Y chromosome as an approach to cloning Y-1inked genes. Using restriction analysis, 150 Y-specific cosmid clones have been screened. Four putative HTF island containing clones were selected and studied in more detail using Southern mapping, northern analysis, library screening, DNA sequencing and methylation analysis. Although data indicate that three of the clones are not associated with transcribed sequences, the fourth clone has been shown through sequence analysis, to contain a possible open reading frame. A direct approach to cloning Hpall tiny fragments from a Y chromosome specific library, obtained from the American Type Culture Collection, is also described. Following the isolation and purification of the human inserts away from the phage vector within this library, the inserts were further restricted using the enzyme Hpall and the resulting tiny fragments re-cloned and analysed for their possible association with transcribed sequences. Although a clone was identified which hybridizes to human genomic sequences, data indicates this to be of autosomal origin. Evidence is also discussed that the human Y chromosome appears to be relatively deficient of HTF islands when compared with the remainder of the genome, and that this deficiency could also reflect the small number of genes which are carried on the human Y chromosome.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available