pHHyDHODH-GFP Expression Plasmid

The pHHyDHODH-GFP expression plasmid is used to express dihydroorotate dehydrogenase (DHODH) fused to green fluorescent protein (GFP) in P. falciparum.

Highlights:

  • Expression of the DHODH-GFP fusion in P. falciparum is driven by a strong HSP86 5’UTR
  • Methotrexate-resistant human DHFR serves as a selectable marker for the plasmid 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.

Catalog Number Product Size AVAILABILITY Price Qty
EDX007
pHHyDHODH-GFP Expression Plasmid
150ng In stock
Price: $66.00
Specifications
Product Type: Plasmid
Gene/insert name: DHODH
Organism: S. cerevisiae (and P. falciparum and Homo sapiens auxiliary DNA elements)
Antibiotic Resistance: Amp (E. coli) / antifolate WR99210 (P. falciparum)
Fusion Tag(s): GFP/myc
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
Promoter: HSP86 5’UTR
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.
Storage: -20C
Shipped: Room Temperature
Provider
From the laboratory of Akhil B. Vaidya, PhD, Drexel University.

References
  1. Cormack B. P., et al. (1996) FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173:33-8.
  2. O'Donnell, R. A. et al. (2002) A genetic screen for improved plasmid segregation reveals a role for Rep20 in the interaction of Plasmodium falciparum chromosomes. EMBO J 21, 1231-9.
  3. Painter HJ, Morrisey JM, Mather MW, Vaidya AB. (2007) Specific role of mitochondrial electron transport in blood-stage Plasmodium falciparum. Nature 446:88-91.

If you publish research with this product, please let us know so we can cite your paper.

 
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