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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_0108300
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Gene Model P. falciparum ortholog |
PF3D7_0609800
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Gene product | palmitoyltransferase DHHC2, putative |
Gene product: Alternative name | DHHC2 |
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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 used | (Linear) plasmid double cross-over |
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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Partial or complete disruption of the gene | Complete |
Additional remarks partial/complete disruption |
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Selectable marker used to select the mutant parasite | tgdhfr |
Promoter of the selectable marker | pbdhfr |
Selection (positive) procedure | pyrimethamine |
Selection (negative) procedure | No |
Additional remarks genetic modification | Attempts to disrupt the dhhc2 gene in P. berghei were unsuccessful indicating an essential role of DHCC2 for blood stage development/multiplication.
Phenotype analyses of promoter-swap mutants indicate that DHHC2 plays an important role in the development of zygotes into mature ookinetes (see mutants RMgm-1349 and RMgm-1351 for more information on the DHHC2 protein).
To disrupt dhhc2 (PBANKA_010830) we constructed a replacement construct which contains the pyrimethamine resistant tgdhfr/ts as a selectable marker cassette flanked by target sequences for homologous recombination; they were PCR-amplified from P. berghei WT genomic DNA using primers 0739 and 0740, and 0741 and 0742 specific for the 5′ or 3′ flanking regions of the dhhc2 ORF, respectively. The final DNA construct pLIS0065 used for transfection was obtained after digestion of the replacement construct with Asp718I and NotI. |
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 | aaaggtaccTTTATTATTTGAGTGTTG |
Additional information primer 1 | g0739 |
Sequence Primer 2 | aaaaagcttTTTATATTGATTTTGATTG |
Additional information primer 2 | g0740c |
Sequence Primer 3 | aaagaattcTTGAGTTTATAATATGTC |
Additional information primer 3 | g0741 |
Sequence Primer 4 | aaagcggccgcATATCCTAAAAACTATTG |
Additional information primer 4 | g0742c |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
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