RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1434
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1222200; Gene model (P.falciparum): PF3D7_0808450; Gene product: protein MPODD, putative (mitochondrial protein ookinete developmental defect (MPODD))
Transgene
 Transgene not Plasmodium: mCherry
Promoter: Gene model: PBANKA_1222200; Gene model (P.falciparum): PF3D7_0808450; Gene product: protein MPODD, putative (mitochondrial protein ookinete developmental defect (MPODD))
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (DHFR-TS)
Replacement locus: Gene model: PBANKA_1222200; Gene product: protein MPODD, putative (mitochondrial protein ookinete developmental defect (MPODD))
Phenotype Fertilization and ookinete; Oocyst;
Last modified: 9 April 2016, 13:22
  *RMgm-1434
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) : 27053680
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone Not applicable
Other information parent line
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The mutant parasite was generated by
Name PI/ResearcherKlug, D; Frischknecht F; Douglas RG
Name Group/DepartmentIntegrative Parasitology, Center for Infectious Diseases
Name InstituteHeidelberg University Medical School
CityHeidelberg
CountryGermany
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Name of the mutant parasite
RMgm numberRMgm-1434
Principal namempodd(-)
Alternative name
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 ookineteAberrant ookinete development. Knockout parasites failed to develop mature, infectious ookinetes. mpodd(-) mutants typically resembled retort forms with the nucleus still positioned in the bulb-like extension of the zygote, suggesting an important role in final conversion from zygotes to ookinetes.
OocystNo oocyst production
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of MPODD.

Protein (function)
Bioinformatic analysis of MPODD revealed the presence of a TM domain as well as a mitochondrial targeting signal.

Phenotype
Normal blood stage development and gametocyte production. Aberrant ookinete development. Knockout parasites failed to develop mature, infectious ookinetes. mpodd(-) mutants typically resembled retort forms with the nucleus still positioned in the bulb-like extension of the zygote, suggesting an important role in final conversion from zygotes to ookinetes. No oocyst production.

Additional information
By analysis of a mutant expressing a mCherry tagged version of MPODD (RMgm-1435) evidence is presented for a location in mitochondria.

Evidence is presented that in the absence of lack of MPODD expression the overall structural integrity of mitochondria was not affected but results in 'reduced mitochondrial mass'.

Evidence is presented that complementation of mpodd(-) parasites with a wildtype P. berghei copy of the mpodd gene rescued the defect in ookinete development and oocyst production

Evidence is presented that complementation of mpodd(-) parasites with the P. falciparum ortholog (PF3D7_0808450) rescued the defect in ookinete development and oocyst production

Other mutants
A mutant expressing a mCherry tagged version of MPODD (RMgm-1435)

  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_1222200
Gene Model P. falciparum ortholog PF3D7_0808450
Gene productprotein MPODD, putative
Gene product: Alternative namemitochondrial protein ookinete developmental defect (MPODD)
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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 modification
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 namemCherry
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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) procedurepyrimethamine
Selection (negative) procedureNo
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_1222200
Gene Model P. falciparum ortholog PF3D7_0808450
Gene productprotein MPODD, putative
Gene product: Alternative namemitochondrial protein ookinete developmental defect (MPODD)
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 / InsertionReplacement locus
Gene Model of Parasite PBANKA_1222200
Gene productprotein MPODD, putative
Gene product: Alternative namemitochondrial protein ookinete developmental defect (MPODD)
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: StudioDrie StudioDrie; S. Mededovic and A. van Wigcheren