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Title: Cloning and characterisation of Drosophila imp dehydrogenase
Author: Slee, Roger B.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1992
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A preliminary molecular characterisation was conducted of a transcription unit located adjacent to an enhancer-trap transposon (p[lac Z, ry±] at 38C, in a transformant exhibiting testis-specific expression of the β-galactosidasereporter gene. Northern analysis with a probe (A-5.1) consisting of a genomic fragment situated immediately upstream of the transposon, identified a region which encoded three transcripts with distinct sizes and expression profiles, including 1.0 kb and 2.5 kb species which are highly enriched in adult ovary RNA. The same probe was used to isolate a cDNA clone (OV.1) representing one of the transcripts detected by A-5.1. In Northern hybridisation OV.1 detected all the same transcripts as A-5.1 plus an additional testis-specific transcript of 0.8 kb. An OV.1 probe was used to isolate a cDNA (T1) corresponding to the testis-specific transcript and T1 was shown to recognise all the same transcripts as OV.1 on a Northern blot except for the 1.0 kb ovary enriched species. Ov.1 and T1 were sequenced and found to represent overlapping transcripts from the same gene. OV.1 contains unique 5' sequences which are encoded immediately upstream of p[lac z, ry±] whilst the region common to OV.1 and T1 is encoded more than 8 kb distant in the genome. The picture that emerged from this analysis was of a gene which included an intron of over 8 kb and generated multiple independently regulated, overlapping transcripts including testis-specific and ovary enriched variants. The conclusion from this research was that the Drosophila IMPD gene has a more complex expression pattern than was predictable on the basis of the enzyme's known functions and molecular data from other species. The possible significance of the observed expression pattern is considered.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available