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Functional expression of the dihydrofolate reductase and thymidylate synthetase activities of the human malaria parasite Plasmodium falciparum in Escherichia coli
- Source :
- Molecular and biochemical parasitology. 45(2)
- Publication Year :
- 1991
-
Abstract
- We have developed a recombinant system that directs the functional expression from Escherichia coli of both dihydrofolate reductase-thymidylate synthetase (DHFR-TS) and the isolated DHFR domain from Plasmodium falciparum. Both products are inhibitory to a number of E. coli cell lines to the extent that cell growth ceases immediately upon induction. This dramatic inhibition is not seen in strain AB1899, in which amounts of plasmodial protein of up to 100 times the basal E. coli TS level can be accumulated. However, as well as the full-length DHFR-TS molecule, smaller proteins carrying an intact TS substrate-binding site are produced. These represent ca. 60-75% of the total plasmodial protein expressed and are observed in every E. coli strain examined. We show that they are not derived by degradation of the parent DHFR-TS molecule, but can be correlated with the sizes of proteins expected to be produced if erroneous initiation of translation were occurring at 3 internal methionine residues.
- Subjects :
- Molecular Sequence Data
Plasmodium falciparum
medicine.disease_cause
Thymidylate synthase
Polymerase Chain Reaction
Gene Expression Regulation, Enzymologic
parasitic diseases
Dihydrofolate reductase
Protein biosynthesis
medicine
Escherichia coli
Animals
Cloning, Molecular
Molecular Biology
biology
Base Sequence
Genetic transfer
Trimethoprim Resistance
Thymidylate Synthase
biology.organism_classification
Enterobacteriaceae
Molecular biology
Recombinant Proteins
Tetrahydrofolate Dehydrogenase
Biochemistry
Protein Biosynthesis
biology.protein
Parasitology
Electrophoresis, Polyacrylamide Gel
Heterologous expression
Plasmids
Subjects
Details
- ISSN :
- 01666851
- Volume :
- 45
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Molecular and biochemical parasitology
- Accession number :
- edsair.doi.dedup.....84c84e3af8aa26c241dfdea3ceb96806