RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
DisruptedGene model (rodent): PBANKA_0214400; Gene model (P.falciparum): PF3D7_0730300; Gene product: AP2 domain transcription factor AP2-L, putative (AP2-L; ApiAP2)
Phenotype Liver stage;
Last modified: 6 February 2013, 09:17
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) : 23144823
MR4 number
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
The mutant parasite was generated by
Name PI/ResearcherS. Iwanaga; M. Yuda
Name Group/DepartmentDepartment of Medical Zoology
Name InstituteMie University School of Medicine
CityTsu, Mie
Name of the mutant parasite
RMgm numberRMgm-792
Principal nameAP2-L(-)
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 stageNormal sporozoite production. Gliding motility, cell traversal and hepatocyte invasion of sporozoites is normal. Infectivity of sporozoites in vivo is 10.000 lower compared to wild type sporozoites (as measured by prepatent period). Development of liver stages is affected (see 'Additional remarks phenotype').
Additional remarks phenotype

The mutant lacks expression of AP2-L (transcription factor with AP2 domain(s))

Protein (function)
Apetala 2 (AP2)-family proteins are transcription factors that have DNA-binding domains of ,60 amino acids called AP2 domains. Recently, AP2 genes have been found in the genomes of Plasmodium parasites. In P. falciparum 27 AP2-family genes have been identified. Among these genes, 26 are conserved in the other Plasmodium species whose entire genomes have been sequenced. Each member of this family has 1 to 4 AP2 domains, and the amino acid sequences of these domains are highly conserved among Plasmodium orthologs.
AP2-L encodes a protein of 1272 amino acids with two AP2 domains. These domains are located near the C-terminus of the protein and are separated from each other by a short linker.

The phenotype analyses indicate that the lack of expression of AP2-L does not affect blood stage and mosquito stage development. Gliding motility, cell traversal and hepatocyte invasion of sporozoites is normal. Liver stage development is strongly reduced.
At 24 hpi, the difference in size between the wild-type and AP2-L(2) liver stages was still subtle, but it became clear at 36 hpi. After 36 hpi, the sizes of the AP2-L(2) LS parasites scarcely increased, whereas the wild-type parasites continued to grow until 48 hpi. In accordance with the growth of the LS parasites, arrest of nuclear division became clear at 36 hpi in the AP2-L(2) parasites. The appearance of the nuclei remained the same, and no AP2-L(2) LS parasites in the cytomere stage were formed in the later stage. These observations indicate that the development of AP2-L(2) parasites almost completely arrested at about 36 hpi, i.e., in the middle of the schizont stage, which caused a severe decrease in their ability to infect the liver

Additional information
Analyses of a mutant expressing a C-terminal tagged version of AP2-L showed expression and localisation of AP2-L in the nucleus of erythrocytic trophozoites, oocyst-derived sporozoites, salivary gland sporozoites and liver stages.

Other mutants
RMgm-793: A mutant expressing a C-terminal GFP-tagged version of AP2-L
RMgm-794: A mutant expressing a C-terminal tagged mCherry version of AP2-L and expressing GFP under a constitutive promoter (eef1a).

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0214400
Gene Model P. falciparum ortholog PF3D7_0730300
Gene productAP2 domain transcription factor AP2-L, putative
Gene product: Alternative nameAP2-L; ApiAP2
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 geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr
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 1AP2-L-1
Additional information primer 2AP2-L-2
Additional information primer 3AP2-L-3
Additional information primer 4AP2-L-4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6