RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-887
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1014500; Gene model (P.falciparum): PF3D7_1430200; Gene product: plasmepsin IX
PhenotypeNo phenotype has been described
Last modified: 8 June 2014, 12:42
  *RMgm-887
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 3
Reference (PubMed-PMID number) Not published (yet)
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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 P. berghei ANKA 507cl1 (RMgm-7)
Other information parent lineP.berghei ANKA 507cl1 (RMgm-7) is a reference ANKA mutant line which expresses GFP under control of a constitutive promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190)
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Attempts to generate the mutant parasite were performed by
Name PI/ResearcherB. Franke-Fayard; J.B. Dame; C.J. Janse
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center (LUMC)
CityLeiden
CountryThe Netherlands
  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_1014500
Gene Model P. falciparum ortholog PF3D7_1430200
Gene productplasmepsin IX
Gene product: Alternative name
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Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid 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 parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe negative attempts to disrupt plasmepsin IX indicates an essential role during blood stage development.

P. berghei has 7 plasmepsins (aspartic proteases (PM IV-PM X). PM IV has a role in hemoglobin digestion (RMgm-314, RMgm-315, RMgm-316). P. falciparum PM V performs a critic role in the endoplasmic reticulum processing the PEXEL sequence of exported proteins. PM VI, VII, VII are not expressed in asexual blood stages; these are expressed in gametocytes/ookinetes/oocysts. PM VI plays an essential role in the formation of sporozoites within the oocyst (RMgm-97).
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: S. Mededovic and A. van Wigcheren