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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_0509500
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Gene Model P. falciparum ortholog |
PF3D7_1025300
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Gene product | conserved Plasmodium protein, unknown function |
Gene product: Alternative name | PSOP21, putative secreted ookinete protein 21 |
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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 | 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 | Partial |
Additional remarks partial/complete disruption |
gene disruption (fragment encoding N-terminal 502(Pf) amino acids retained)
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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 | IMPORTANT NOTE: The new P. berghei annotation, in GeneDB, has revealed that the primers for the (5’ and 3’) targeting regions are located in TWO different genes (PBANKA_050950 and PBANKA_050940). Therefore the integration of this construct would target both genes, i.e. generating a partial deletion of PBANKA_050950 and a complete deletion of PBANKA_050940. The failure to disrupt/knock-out PBANKA-050950 might, therefore, be due to the unintentional disruption of PBANKA_050940. |
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 | TTGGGCCCCAAAGGGAATATGGAATATTTAG |
Additional information primer 1 | AE21a (ApaI); 5' |
Sequence Primer 2 | CCAAGCTTACATCATCAGTATTTTTACAATC |
Additional information primer 2 | AE21b (HindIII); 5' |
Sequence Primer 3 | TGAATTCATATTAATAAGTTAGGCGAAACC |
Additional information primer 3 | AE21c (EcoRI); 3' |
Sequence Primer 4 | GGGGATCCTGCACATATTATATGCTTAACTG |
Additional information primer 4 | AE21d (BamHI); 3' |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
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