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Characterisation and expression of the carbamate kinase gene from Giardia intestinalis.
- Source :
-
Molecular and biochemical parasitology [Mol Biochem Parasitol] 1999 Jan 05; Vol. 98 (1), pp. 43-51. - Publication Year :
- 1999
-
Abstract
- The arginine dihydrolase pathway in Giardia intestinalis produces energy via the carbamate kinase (CBK, ATP:carbamate phosphotransferase, EC 2.7.2.2) reaction. Characterisation of the CBK gene from the Portland 1 strain indicated that it is located on either chromosome 3 or 4, does not appear to contain introns and is expressed in both the trophozoite and early cyst stages. Heterologous expression of CBK in Escherichia coli, using the pQE-30 expression system (QIAGEN), enabled a one-step purification of the recombinant enzyme via affinity chromatography. The expressed protein was identified by enzyme assay and mass spectrometry. The native and recombinant forms of the enzyme have similar physical properties and the recombinant enzyme appears to be active as the homodimer.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Chromosome Mapping
Cloning, Molecular
Energy Metabolism
Giardia lamblia enzymology
Hydrolases
Molecular Sequence Data
Molecular Weight
Phosphotransferases (Carboxyl Group Acceptor) biosynthesis
Phosphotransferases (Carboxyl Group Acceptor) isolation & purification
Recombinant Proteins biosynthesis
Recombinant Proteins isolation & purification
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Genes, Protozoan
Giardia lamblia genetics
Phosphotransferases (Carboxyl Group Acceptor) genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0166-6851
- Volume :
- 98
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Molecular and biochemical parasitology
- Publication Type :
- Academic Journal
- Accession number :
- 10029308
- Full Text :
- https://doi.org/10.1016/s0166-6851(98)00141-8