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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_1410950
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Gene Model P. falciparum ortholog |
PF3D7_1312450
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Gene product | apical ring associated protein 1, putative |
Gene product: Alternative name | ARA1 |
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Details of the genetic modification |
Name of the tag | GFP |
Details of tagging | C-terminal |
Additional remarks: tagging | To examine localisation of the protein, the ORF was expressed as a GFP-tagged protein under control of the original promoter, using the centromere plasmid pCen-GFP. |
Commercial source of tag-antibodies | |
Type of plasmid/construct | Plasmodium Artificial Chromosome (PAC) circulair |
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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Selectable marker used to select the mutant parasite | tgdhfr |
Promoter of the selectable marker | eef1a |
Selection (positive) procedure | pyrimethamine |
Selection (negative) procedure | No |
Additional remarks genetic modification | Protein targeting was investigated by expressing the gene as a GFP-tagged protein, using the centromere plasmid pCen-GFP (circular PAC). Target genes with the upstream regulatory region (1.1–1.5-kbp upstream of the first methionine codon) were amplified by PCR using genomic DNA as the template, and then subcloned into the region immediately upstream of the GFP gene of the pCen vector. |
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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