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Title: Characterisation of newly isolated genes expressed in the ovary of Drosophila melanogaster
Author: Grimes, Brenda R.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1990
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Two genomic clones called λov5 and λov2 were isolated from a differential screen and found to encode ovary enriched transcripts. The genes within λov2 and λov5 have been initially characterised with a view to ultimately determining their function(s) during oogenesis. λ0v5 consists of two EcoRI sub-fragments denoted ov5A and ov5B. Each of these fragments detect ovary enriched transcripts on Northern blots. The ov5A probe was used to screen a λgt11 ovary cDNA library and three size classes of cDNA called λov5A1, λov5A2 and λov5A5 were identified. Each cDNA detects at least two ovary enriched transcripts, approximately 1.7 kb and 1.4 kb in length. The ov5A5 cDNA also hybridises with two additional ovary specific transcripts approximately 0.8 kb and 0.2 kb in length. The organisation of these ovary transcripts within the ov5A genomic region was determined by cross-hybridisation of cDNAs to Southern blots of restriction enzyme digests of ov5A sequences. The ov5B sub-fragment detects an ovary specific transcript approximately 1.4 kb in length. Full length cDNA copies of this transcript were isolated. In situ hybridisation to whole mounts of Drosophila ovaries were carried out using digoxigenin-labelled ov5A2 and ov5B1 cDNA probes to determine the spatial and temporal distribution of the ovary enriched transcripts during oogenesis. ov5A2 detects transcripts in the nurse cell cytoplasm and in oocytes from stage 7 onwards. ov5B1 strongly detects transcripts in stage 10B nurse cell cytoplasm and in the degenerating nurse cells. Two EcoRI sub-fragments of λov2, ov2A and ov2B, hybridise with female cDNA but not with male cDNA. ov2A detects two ovary and embryo specific transcripts approximately 4.0 kb and 3.0 kb in size and also a 1.6 kb sized male-specific transcript. ov2B detects two transcripts estimated to be 3.5 kb and 2.5 kb in length which are expressed exclusively in the ovary. λov5 and λov2 were mapped on chromosome 3R at positions 88B/C and 89B respectively. Within the cytological region 88-89, two female sterile mutations called Spindle-B and fs293gamma5 have been genetically mapped. Experiments with a set of deficiency chromosomes were carried out to determine whether or not the ov5 or ov2 genomic sequences mapped close to these mutations.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available