RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-188
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_0408500; Gene model (P.falciparum): PF3D7_0310400; Gene product: parasite-infected erythrocyte surface protein | TVN-junction protein 1
PhenotypeNo phenotype has been described
Last modified: 16 March 2010, 18:27
  *RMgm-188
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 2
Reference (PubMed-PMID number) Reference 1 (PMID number) : 18551176
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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 cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
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Attempts to generate the mutant parasite were performed by
Name PI/ResearcherC. van Ooij; C.J. Janse; K. Haldar
Name Group/DepartmentDepartment of Pathology
Name InstituteNorthwestern University
CityChicago, Illinois
CountryUSA
  Disrupted: Mutant parasite with a disrupted gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0408500
Gene Model P. falciparum ortholog PF3D7_0310400
Gene productparasite-infected erythrocyte surface protein | TVN-junction protein 1
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 KpnI/SacII
Partial or complete disruption of the geneComplete
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 gene has been identified in a study to identify proteins that are exported from the blood stage parasite to the host erythrocyte and that are conserved between P. falciparum and P. berghei. These proteins contain signal sequences and the Host-Targeting (HT) motif (van Ooij et al., 2008, PloS Pathogens 4, e10000084). Eleven conserved proteins were identified and 9 genes encoding these proteins were targeted for gene disruption in P. berghei. All attempts to disrupt these genes were unsuccessful, indication the essential nature of their proteins for blood stage development.

Name of the transfection attempts experiments (Leiden Malaria Research Group): 712.6; 719.4
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 1AAAAAGCAGGCTccgcggCCGAGAATGTATATATACTAAACGTGCTTAGCT
Additional information primer 1PB001106.03.0 upstream (SacII)
Sequence Primer 2AGAAAGCTGGGTactagtCTTTCGCTTTTTTGCATGAAATAAATCACTGTG
Additional information primer 2PB001106.03.0 upstream (SpeI)
Sequence Primer 3AAAAAGCAGGCTaagcttCCAATAACTAatTGTGttACTTAttctGGGCGTTC
Additional information primer 3PB001106.03.0 downstream (HindIII)
Sequence Primer 4AGAAAGCTGGGTggtaccTGGACTATAATTATATTTTTTCAAGTTATGTATTACATCCAG
Additional information primer 4PB001106.03.0 downstream (KpnI)
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