RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-1431
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_0207000; Gene model (P.falciparum): PF3D7_0106300; Gene product: calcium-transporting ATPase (ATP6; SERCA; ATPase6)
PhenotypeNo phenotype has been described
Last modified: 30 March 2016, 10:54
  *RMgm-1431
Successful modificationThe gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted Gene disruption
Number of attempts to introduce the genetic modification Unknown
Reference (PubMed-PMID number) Reference 1 (PMID number) : 23599312
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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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Attempts to generate the mutant parasite were performed by
Name PI/ResearcherPulcini S; Tewari R; Krishna S
Name Group/DepartmentDivision of Clinical Sciences
Name InstituteSt. George's, University of London
CityLondon
CountryUK
  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0207000
Gene Model P. falciparum ortholog PF3D7_0106300
Gene productcalcium-transporting ATPase
Gene product: Alternative nameATP6; SERCA; ATPase6
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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 geneUnknown
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasiteunknown
Promoter of the selectable markerunknown
Selection (positive) procedureunknown
Selection (negative) procedureNo
Additional remarks genetic modificationUnsuccessful attempts to disrupt or tag this gene indicates an essential role of ATP6 (SERCA) for growth/multiplication of blood stages.

Limited information is provided in the paper on the genetic modification attempts: "Similarly, a double recombination construct failed to KO pbserca in the more tractable Plasmodium berghei system, and C-terminal tagging was also unsuccessful (data not shown).

Also attempts to disrupt/tag the P. falciparum ortholog were unsuccessful.

ATP6 is the only SERCA-type Ca2+-ATPase sequence in the Plasmodium genome. The protein is thought to be a P-type ATPase involved in calcium ion transport. Mutations in this gene has been related to artemisinin-resistance of malaria parasites but results from several studies indicate that PfATP6 is not directly involved in artemisinin action or resistance.
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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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