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| Details of the target gene |
| Gene Model of Rodent Parasite |
PBANKA_0105900
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| Gene Model P. falciparum ortholog |
PF3D7_1440300
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| Gene product | delta-aminolevulinic acid dehydratase | porphobilinogen synthase |
| Gene product: Alternative name | PBGS; ALAD |
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| Details of the genetic modification |
| Inducable system used | No |
| Additional remarks inducable system |
|
| Type of plasmid/construct used | (Linear) plasmid double cross-over |
| PlasmoGEM (Sanger) construct/vector used | No |
| Modified PlasmoGEM construct/vector used | No
|
| Plasmid/construct map |
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| Plasmid/construct sequence |
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| Restriction sites to linearize plasmid |
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| Partial or complete disruption of the gene | Complete |
| Additional remarks partial/complete disruption |
|
| Selectable marker used to select the mutant parasite | hdhfr/yfcu |
| Promoter of the selectable marker | pbdhfr |
| Selection (positive) procedure | pyrimethamine |
| Selection (negative) procedure | No |
| Additional remarks genetic modification | Unsuccessful attempts to disrupt the gene indicate that this gene is essential for asexual blood stage growth/multiplication.
Fragments of the 3’ and 5’ UTR were amplified from gDNA using gene-specific primers. First, 3’ fragments were cloned into the pBAT vector, following restriction digestion with HindIII and KpnI to generate an intermediate construct (pHEME-im). Then, the 5’ UTR fragment was digested with SacII and EcoRV and cloned into the respective SacII- and PvuII-linearized pHEME-im construct, thus removing the mCherry-3xMyc tag. Finally, the pHEME-ko vector was linearized with ScaI and SalI before transfection into P. berghei strain ANKA (wt) parasites. |
| Additional remarks selection procedure | |
Primer information: Primers used for amplification of the target sequences  Primer information: 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 | |
| Sequence Primer 5 | |
| Additional information primer 5 | |
| Sequence Primer 6 | |
| Additional information primer 6 | |
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*RMgm-1587