RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-4109
Malaria parasiteP. yoelii
Genotype
DisruptedGene model (rodent): PY17X_1126500; Gene model (P.falciparum): PF3D7_0626300; Gene product: 3-oxoacyl-acyl-carrier protein synthase I/II (FABB/F)
Transgene
 Transgene not Plasmodium: GFP-Luciferase
Promoter: Gene model: PBANKA_1133300; Gene model (P.falciparum): PF3D7_1357100; Gene product: elongation factor 1-alpha (eef1a)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Insertion locus: Gene model: PY17X_0306600; Gene product: 6-cysteine protein (P230p; 230p)
Phenotype Liver stage;
Last modified: 21 January 2017, 14:15
  *RMgm-4109
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption, Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 22216235
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. yoelii
Parent strain/lineP. y. yoelii 17XNL
Name parent line/clone RMgm-689
Other information parent lineRMgm-689 is a reference P. yoelii 17XNL line (1971cl1) that expresses the fusion protein GFP-Luciferase under the control of the constitutive P. berghei eef1a promoter. The transgene expression cassette is introduced into the 230p locus. This line does not contain a drug-selectable marker.
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The mutant parasite was generated by
Name PI/ResearcherC.J. Janse; S.M. Khan, B.M. Franke-Fayard
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
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Name of the mutant parasite
RMgm numberRMgm-4109
Principal name2251cl3
Alternative nameΔPyFabBF-GFP-Luc(con)
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageGrowth arrest of late liver stages
Additional remarks phenotype

Mutant/mutation
The mutant lack expression of FABB/F and expresses the fusion protein GFP-Luciferase under the control of the constitutive P. berghei eef1a promoter.

Phenotype
Late arrest of liver stages. See also the detailed phenotype analyses of a P. yoelli mutant RMgm-183 that lacks expression of FABB/F

Additional information

Other mutants

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PY17X_1126500
Gene Model P. falciparum ortholog PF3D7_0626300
Gene product3-oxoacyl-acyl-carrier protein synthase I/II
Gene product: Alternative nameFABB/F
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThis mutant was generated in the reference line 1971cl1 (see above) by standard methods of transfection using a DNA construct that targets the fabb/f gene by double cross-over integration. The 5’- and 3’- fabb/f target regions were amplified using primers 7358 5’-atgggcccTTGCGCTATTTATAAGAGTTTGAGAGG / 7359 5’-aaggcctCAAGAATATTTTTAAGGGCCATTTC and 7360 5’-ccggggtaccCAATGATTGCAAATACACCATCAG / 7361 5’-ataagaatgcggccgcGTGGATATACGCAAGTGTGCGAG and cloned up (ApaI/StuI) and downstream (KpnI/NotI) of the hdhfr/fcu selectable marker cassette of plasmid pL0034. The final DNA construct was linearized with HindIII/EcoRI before transfection.
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
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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  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFP-Luciferase
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedureNo
Selection (negative) procedure5-fluorocytosine (5-FC)
Additional remarks genetic modification
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 productelongation factor 1-alpha
Gene product: Alternative nameeef1a
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
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 productbifunctional dihydrofolate reductase-thymidylate synthase, putative
Gene product: Alternative namedhfr/ts
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionInsertion locus
Gene Model of Parasite PY17X_0306600
Gene product6-cysteine protein
Gene product: Alternative nameP230p; 230p
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the primers used for amplification of the target sequences  Click to hide information
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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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren