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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_1129000
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Gene Model P. falciparum ortholog |
PF3D7_0630200
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Gene product | secreted ookinete protein, putative |
Gene product: Alternative name | PSOP6, putative secreted ookinete protein 6 |
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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 | 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 | The PSOP6 protein is detected in a proteome analysis of ookinetes.
Integration of the gene targeting construct into the genomic locus was detected in some transfections by diagnostic PCR, suggesting that these genes are not essential in the asexual blood stages. However, the mutants were never abundant enough in the mixed drug-resistant populations which also contains episome-containing wild type parasites) to allow cloning. It is unclear whether this is due to technical reasons or a reduced growth rate of the mutants. |
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 | TTGGGCCCGGATCTTTGTAACATTTGGCATG |
Additional information primer 1 | AE06a (ApaI), |
Sequence Primer 2 | CCAAGCTTTTTATATCCACACAGTGTAAGAG |
Additional information primer 2 | AE06b (HindIII), |
Sequence Primer 3 | TGAATTCAGTAAAATGCACACACAACATTC |
Additional information primer 3 | AE06c (EcoRI), |
Sequence Primer 4 | GGGGATCCTTGGCATAACATATAATTTCACG |
Additional information primer 4 | AE06d (BamHI), |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
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