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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_0302500
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Gene Model P. falciparum ortholog |
PF3D7_0204700
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Gene product | hexose transporter |
Gene product: Alternative name | hexose transporter 1, HT1; PfHT1, PfHT, PbHT, PbHT1 |
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Details of the genetic modification |
Name of the tag | HA-tag |
Details of tagging | C-terminal |
Additional remarks: tagging | The endogenous pbht1 gene is replaced with a HA-tagged copy of pbht1. |
Commercial source of tag-antibodies | |
Type of plasmid/construct | 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 |
KpnI, NotI
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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 endogenous pbht1 gene is replaced with a HA-tagged copy of pbht1. Primers 1-4 show the primers used to amplify the 3'- and 5'-target regions of pbht1. Primer 5-6 are the primers to generate/amplify the HA-tagged form of pbht1 |
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 | CTCATCGCGGCCGCGCAAAACTAAATCGGATTAATGCCGA |
Additional information primer 1 | PbHT1–5’UTR-F, NotI |
Sequence Primer 2 | CTCATCGGATCCACGCAATATATTCATTTTTTCGTATTAATACACATATATTTCTTG |
Additional information primer 2 | PbHT1–5’UTR-R, BamHI |
Sequence Primer 3 | CTCATCGGGCCCCATACAAGAACACACCGGCAAT |
Additional information primer 3 | PbHT1–3’UTR-F, ApaI |
Sequence Primer 4 | CTCATCGGTACCCTTATGTTAAACAATTATCCTTTCCAATTATCACAC |
Additional information primer 4 | PbHT1–3’UTR-R, KpnI |
Sequence Primer 5 | CTCATCGCGGCCGCGCAAAACTAAATCGGATTAATGCCGA |
Additional information primer 5 | PbHT1–5’UTR-F, NotI |
Sequence Primer 6 | CTCATCGGATCCTTAAGCGTAATCTGGAACATCGTATGGGTAAACTCTTGATTTGCTTATATGTTTTTGTCTTTCTTC |
Additional information primer 6 | PbHT1-HA-R, BamHI |
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