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Type and details of transgene |
Is the transgene Plasmodium derived |
Transgene: not Plasmodium |
Transgene name | GFP |
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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 | PCR construct 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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Selectable marker used to select the mutant parasite | hdhfr/yfcu |
Promoter of the selectable marker | pbdhfr |
Selection (positive) procedure | pyrimethamine |
Selection (negative) procedure | No |
Additional remarks genetic modification | The construct to tag the nfuapi gene contains a GFP expression cassette. In this cassette the gfp gene is under the control of the hsp70 promoter (see for more information on the basic plasmid pBAT-SIL6 the mutant RMgm-757).
PCR fragments were cloned into the berghei adaptable transfection vector pBAT-SIL6. First, the 3'UTR homologous sequences were cloned following restriction digestion of vector and insert with HindIII and KpnI (see RMgm-909).
Next the carboxy-terminal part of NFUapi was PCR amplified using gene-specific primers. After restriction digestion with SacII and EcoRV, the fragment was cloned into the SacII and HpaI digested pBAT-SIL6 vector already containing the 3'UTR sequence of NFUapi, thus fusing the NFUapi carboxyterminal sequence in frame with the mCherry-3xMyc tag sequence. The resulting plasmid was linearized with SalI and used to transfect P. berghei strain ANKA (WT) parasites. |
Additional remarks selection procedure | Cloned by FACS sorting of GFP-expressing parasites |
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Other details transgene |
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Promoter |
Gene Model of Parasite |
PBANKA_0711900
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Gene Model P. falciparum ortholog |
PF3D7_0818900
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Gene product | heat shock protein 70 |
Gene product: Alternative name | HSP70 |
Primer information details of the primers used for amplification of the promoter sequence
Primer information details of the primers used for amplification of the promoter sequence
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
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3'-UTR |
Gene Model of Parasite |
PBANKA_1340000
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Gene product | dihydrofolate synthase/folylpolyglutamate synthase, putative |
Gene product: Alternative name | PbDHFS-FPGS |
Primer information details of the primers used for amplification the 3'-UTR sequences
Primer information details of the primers used for amplification the 3'-UTR sequences
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
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Insertion/Replacement locus |
Replacement / Insertion | Replacement locus |
Gene Model of Parasite |
PBANKA_0822300
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Gene product | NifU-like scaffold protein |
Gene product: Alternative name | NifU; NFUapi |
Primer information details of the primers used for amplification of the target sequences
Primer information details of the 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 | |
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