Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.649672
Title: The cloning and characterisation of the genes encoding replication factor C subunits from the malarial parasite, Plasmodium falciparum
Author: Douglas, Jill Karen
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1999
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Abstract:
A better understanding of the molecular biology of the essential cellular processes in Plasmodium falciparum is required if new drug targets are to be discovered against malaria. PfRFC1 is a single copy gene present on chromosome 2. It has an open reading frame of 2712bp, which predicts a protein of 904 amino acids with a molecular weight of 104kDa. It has a transcript of 4kb. PfRFC2 is also present as a single copy gene on chromosome 2. It has an open reading frame of 990bp, which predicts a protein of 330 amino acids with a molecular weight of 38kDa. It has a transcript of 1.6kb. PfRFC3 is present as a single copy on chromosome 14. It has an open reading frame of 1032bp, which predicts a protein of 344 amino acids with a molecular weight of 39kDa. There is one intron of 250bp present at the 5' end of the gene. It has two transcripts of 1.4 and 1.8kb. Small fragments of the three genes were expressed as histidine fusion proteins in E. coli; these were used to make polyclonal antisera in rabbits. Fully-length expression of both PfRfc1 and PfRfc2 was attempted in E. coli both as histidine and GST fusion proteins. However, only the expression of PfRfc2 was successful. Expression of the three genes has been followed during the intraerythrocytic stages of the parasites lifecycle. Northern analysis showed that the transcripts of all three accumulate in trophozoite and schizont stages. Interestingly, PfRFC2 has two larger transcripts of 2.5 and 4kb only present in the schizont sample. The antisera raised against the three genes were used in western analysis and immunofluorescence assays. A similar pattern was seen here with the proteins accumulating in the trophozoite and schizont stages. Anti-RfRfc2 recognised two bands of approximately 100kDa while anti-PfRfc2 and anti-RfRfc3 both recognised proteins of approximately 32kDa. The immunofluorescence assays showed that the proteins are localised in the nucleus.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.649672  DOI: Not available
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