RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
DisruptedGene model (rodent): PBANKA_1013300; Gene model (P.falciparum): PF3D7_1431500; Gene product: mitogen-activated protein kinase 1 (map-1; MAP1; MAPK1)
PhenotypeNo phenotype has been described
Last modified: 2 April 2013, 11:31
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 20951971
Reference 2 (PMID number) : 23544094
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 2.34
Other information parent lineP. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
The mutant parasite was generated by
Name PI/ResearcherR. Tewari, O. Billker
Name Group/DepartmentUniversity of Nottingham
Name InstituteUniversity of Nottingham
Name of the mutant parasite
RMgm numberRMgm-526
Principal nameK11 map-1
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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 stageNot different from wild type
Additional remarks phenotype

The gene has been targeted for gene deletion using a construct aimed at integration into the genome by double cross-over homologous recombination

The gene has been targeted for disruption in a 'kinome-wide' study for deletion of genes encoding Plasmodium protein kinases (protein kinase-like proteins).

See the paper for additional information on the analysis of the phenotype.

See also the paper by Dorin-Semblat D. et al. (2007, Mol Microbiol 65, 1170-1180) for  map-1 knock-out mutants in P. falciparum. Phenotype analyses of these mutants indicate that Pfmap-1, is required neither for in vitro schizogony and gametocytogenesis, nor for gametogenesis and sporogony in the mosquito vector.

Disruption of the P. falciparum ortholog has also been attempted bySolyakov et al. (2011, Nat Commun, 2:565). After transfection with a KO vector a strong PCR signal diagnostic for gene disruption was observed in transfected populations indicating that this gene is not essential for asexual proliferation.

The P. berghei mutant described here has been analysed in more detail for liver stage development by Wierk et al. (2013, PloS One, 8(3): e59755; PMID: 23544094). Development of mutant liver stages was not different from that of wild type liver stages. In the same study they analysed a P. berghei mutant expressing a GFP-tagged version of MAP1 (see RMgm-855) and observed the following: Live cell imaging of transgenic parasites expressing GFP-tagged PbMAPK1 revealed a nuclear localization of PbMAPK1 in the early schizont stage mediated by nuclear localization signals in the C-terminal domain.

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1013300
Gene Model P. falciparum ortholog PF3D7_1431500
Gene productmitogen-activated protein kinase 1
Gene product: Alternative namemap-1; MAP1; MAPK1
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 genePartial
Additional remarks partial/complete disruption nearly complete deletion, including kinase domain
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
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
Additional information primer 1primer sequence target region 1a
Additional information primer 2primer sequence target region 1b
Additional information primer 3primer sequence target region 2a
Additional information primer 4primer sequence target region 2b
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6