Cloning and heterologous expression of Plasmodium ovale dihydrofolate reductase-thymidylate synthase gene
Under a Creative Commons license
open access
Graphical abstract
Highlights
► Polymorphism of P. ovale dihydrofolate reductase-thymidylate synthase was identified. ► Bacterial complementation assay revealed the function of the putative PoDHFR-TS. ► The protein was expressed and purified. ► Biochemical and kinetic properties including antifolate sensitivity were determined.
Abbreviations
DHFR
dihydrofolate reductase
TS
thymidylate synthase
JR
junction region
Pf
Plasmodium falciparum
Po
P. ovale
Pv
P. vivax
Tc
Trypanosoma cruzi
CYC
cycloguanil
DTT
dithiothreitol
dUMP
deoxyuridine monophosphate
EDTA
ethylenediaminetetraacetic acid
HCOH
formaldehyde
H2folate
7,8-dihydrofolate
IPTG
iso-propyl-beta-d-thio-galactopyranoside
LC-MS/MS
liquid chromatography-mass spectrometry
MTX
methotrexate
NADPH
nicotinamide adenine dinucleotide phosphate
PVDF
polyvinylidene fluoride
PYR
pyrimethamine
TES
N-Tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid
TMP
trimethoprim
6-[3H] FdUMP
5-fluoro 2′-deoxyuridine 5′-monophosphate
(6R)-CH2H4folate
5,10-methylene tetrahydrofolate
MM
minimum media
Keywords
Plasmodium ovale
Dihydrofolate reductase-thymidylate synthase
Antifolates
Cited by (0)
Copyright © 2011 Elsevier Ireland Ltd.