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Type and details of transgene |
Is the transgene Plasmodium derived |
Transgene: not Plasmodium |
Transgene name | OVA (ovalbumin) |
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Details of the genetic modification |
Inducable system used | No |
Additional remarks inducable system |
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Type of plasmid/construct | Plasmodium Artificial Chromosome (PAC) circulair |
PlasmoGEM (Sanger) construct/vector used | No |
Modified PlasmoGEM construct/vector used | No
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Plasmid/construct map |
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Plasmid/construct sequence |
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Restriction sites to linearize plasmid |
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Selectable marker used to select the mutant parasite | pbdhfr |
Promoter of the selectable marker | eef1a |
Selection (positive) procedure | pyrimethamine |
Selection (negative) procedure | No |
Additional remarks genetic modification | PyNL-GFP-OVA was generated using a PAC as described by Iwanaga et al (2010, Cell Host & Microbe 7, 245-255) . The following elements were inserted into the PAC: 1) elongation factor promoter; 2) green fluorescence protein (GFP); 3) Pb hsp70 3'-untranslated region; 4) P. berghei (Pb) pyrimethamine resistant gene dihydrofolate reductase-thymidyltransferase-ts (DHFR-ts) gene; 5) Pb DHFR-ts 3'-untranslated region and 6) GFP-OVA, cytosolic form without the ovalbumin (OVA) signal sequence open reading frame.
In the paper no details are provided on (the generation of) the PAC such as centromeric (and telomeric regions; P. berghei or P. yoelii sequences?); circular or linear PAC. In addition no primer sequences (and gene IDs) are provided for amplification of the different regulatory sequences used or for OVA.
The PAC used has been introduced as a circular PAC containing the P. yoelii chromosome 5 centromeric region. |
Additional remarks selection procedure | |
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Other details transgene |
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Promoter |
Gene Model of Parasite |
Not available
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Gene Model P. falciparum ortholog |
Not available
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Gene product | elongation factor 1-alpha |
Gene product: Alternative name | eef1a |
Primer information details of the primers used for amplification of the promoter sequence
Primer information details of the primers used for amplification of the promoter sequence
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
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3'-UTR |
Gene Model of Parasite |
PBANKA_0711900
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Gene product | heat shock protein 70 |
Gene product: Alternative name | HSP70 |
Primer information details of the primers used for amplification the 3'-UTR sequences
Primer information details of the primers used for amplification the 3'-UTR sequences
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
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Insertion/Replacement locus |
Replacement / Insertion | Not available |
Gene Model of Parasite |
Not available
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Gene product | Not available |
Gene product: Alternative name | |
Primer information details of the primers used for amplification of the target sequences
Primer information details of the primers used for amplification of the target sequences
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
Sequence Primer 3 | |
Additional information primer 3 | |
Sequence Primer 4 | |
Additional information primer 4 | |
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