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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_0309500
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Gene Model P. falciparum ortholog |
PF3D7_0212600
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Gene product | secreted protein with altered thrombospondin repeat domain |
Gene product: Alternative name | SPATR |
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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 | Yes |
Name of PlasmoGEM construct/vector | PbGEM-270402 |
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 | hdhfr/yfcu |
Promoter of the selectable marker | eef1a |
Selection (positive) procedure | pyrimethamine |
Selection (negative) procedure | No |
Additional remarks genetic modification | The unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood-stage growth/multiplication.
Deletion of the majority of the spatr gene (PBANKA_0309500) by double cross-over homologous recombination was attempted using the NotI-linearized PbGEM-270402 vector (PlasmoGEM, Sanger). A second construct for total gene deletion was generated by cloning the 5’ (843 bp) and 3’ (292 bp) homology regions amplified by PCR using a Taq DNA polymerase with proofreading activity (Takara) and primers P9 + P12 and P13 + P14 into the pL0001 vector (BEI Resources) via the KpnI/ClaI and EcoRI/BamHI cloning sites, respectively. The linearized constructs were used to transfect synchronized schizonts with the Nucleofector® device (Amaxa). |
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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