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Type and details of transgene |
Is the transgene Plasmodium derived |
Transgene: Plasmodium |
Gene Model of Parasite |
PY17X_1446500
|
Gene Model P. falciparum ortholog |
PF3D7_1229400
|
Gene product | macrophage migration inhibitory factor |
Gene product: Alternative name | MIF |
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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 | 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, SacII
|
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 complete coding region of pymif was PCR amplified from the cDNA clone MS36 obtained from a sequenced P. yoelii 17XL blood-stage cDNA library using forward 5'-GATATCATATGATGCCTTGCTGCGAATTAATAACAAAC-3´ and reverse 5´-GAATTCTCGAGTTAGCCAAATAGTGAACCACTAAAA–3´ primers.
A BamHI pymif cDNA fragment was cloned into gel-purified pPbGFPcon after excision of the PY00068 gene to generate pPyMIF(dhfr). The pPY00068-OE plasmid contains a gfp expression cassette under the control of a constitutive P. berghei eef1a promoter, designed to replace the pydhfr locus by a double crossover event.
Genetic analysis of Py17XL-MIF(+) parasites showed the presence of an intact, endogenous copy of the pymif gene and the integration of the Pb-eef1a-pymif construct at the 5’ end of the pydhfr gene locus via a single crossover event instead of the expected double crossover. |
Additional remarks selection procedure | |
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Other details transgene |
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Promoter |
Gene Model of Parasite |
PBANKA_1133300
|
Gene Model P. falciparum ortholog |
PF3D7_1357100
|
Gene product | elongation factor 1-alpha |
Gene product: Alternative name | eef1a |
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_0719300
|
Gene product | bifunctional dihydrofolate reductase-thymidylate synthase, putative |
Gene product: Alternative name | dhfr/ts |
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 |
PY04370
|
Gene product | bifunctional dihydrofolate reductase-thymidylate synthase |
Gene product: Alternative name | dhfr/ts |
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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