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Heterologous expression of CTP:phosphocholine cytidylyltransferase from Plasmodium falciparum rescues Chinese Hamster Ovary cells deficient in the Kennedy phosphatidylcholine biosynthesis pathway

Sci Rep. 2018-06; 
Marton L,, Hajdú F,, Nagy GN,,, Kucsma N, Szakács G,, Vértessy BG
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Bacterial Expression System The PfCCT 3D7 cDNA sequence (PlasmoDB: PF3D7_1316600) was codon-optimized for expression in E. coli (GenScript, Piscataway, NJ, USA)...resynthesis of the previously codon-optimized cDNA sequence was necessary (GenScript, Piscataway, NJ, USA) to install unique restriction sites for cloning and mutagenesis (for details see Supplementary Information). Get A Quote

摘要

The plasmodial CTP:phosphocholine cytidylyltransferase (PfCCT) is a promising antimalarial target, which can be inhibited to exploit the need for increased lipid biosynthesis during the erythrocytic life stage of Plasmodium falciparum. Notable structural and regulatory differences of plasmodial and mammalian CCTs offer the possibility to develop species-specific inhibitors. The aim of this study was to use CHO-MT58 cells expressing a temperature-sensitive mutant CCT for the functional characterization of PfCCT. We show that heterologous expression of wild type PfCCT restores the viability of CHO-MT58 cells at non-permissive (40 °C) temperatures, whereas catalytically perturbed or structurally destabilized Pf... More

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