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Details of the target gene |
Gene Model of Rodent Parasite |
PCHAS_0101300
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Gene Model P. falciparum ortholog |
Not available
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Gene product | schizont membrane associated cytoadherence protein, putative |
Gene product: Alternative name | SMAC |
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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 | Plasmodium chabaudi AS parasites lacking expression of SMAC (PcASΔsmac) were generated by replacing the P. berghei smac (PBANKA_0100600) targeting region with the equivalent SMAC region of P. chabaudi (PCHAS_0101300; PCHAS_01_v3 52494-53200 and 53700-54284) using a published construct.
Δsmac parasites expressing luciferase constitutively were subsequently generated using a modified construct (PcASΔsmacEFluc; see RMgm-4140) whereby eef1a luciferase was inserted within the targeting region at EcoRV/EcoR1 sites. Both Δsmac and ΔsmacEFluc constructs were digested with Kpn1/SacII prior to transfection. |
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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