The pHHyDHODH-GFP expression plasmid is used to express dihydroorotate dehydrogenase (DHODH) fused to green fluorescent protein (GFP) in P. falciparum.
Yeast DHODH and Aequorea GFPmut2 (1) coding sequences were inserted into an E. coli-Plasmodium falciparum shuttle plasmid (pHHMC*/3R0.5) (2), such that DHODH-GFP were fused into a single coding sequence (including a 10Ala-1Gly linker segment).
From the laboratory of Akhil B. Vaidya, PhD, Drexel University.
|Organism:||S. cerevisiae (and P. falciparum and Homo sapiens auxiliary DNA elements)|
|Antibiotic Resistance:||Amp (E. coli) / antifolate WR99210 (P. falciparum)|
|Grow in E. coli at 37 C:||Yes|
|Selectable markers:||bla, DHFRmtxR, DHODH|
|Cloning Site 5':||BstBI|
|Cloning Site 3':||XhoI and PstI|
|Insert Size:||942 bp|
|Vector Backbone and Size:||6158 bp|
|High or low copy:||High|
|Comments:||Note that the DHODH and GFP sequences are native sequences, i.e., not codon-optimized for either malaria or human expression. DHODH can also serve as a selectable marker by using specific inhibitors of P. falciparum DHODH or of cytochrome bc1.|
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