Summary

RMgm-1495
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1449000; Gene model (P.falciparum): PF3D7_1234400; Gene product: aspartyl protease, putative
PhenotypeNo phenotype has been described
Last modified: 11 July 2016, 18:02
  *RMgm-1495
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) : 27371728
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/ResearcherKehrer J; Frischknecht F, Mair GR
Name Group/DepartmentUniversity of Heidelberg Medical School
Name InstituteUniversity of Heidelberg Medical School
CityHeidelberg
CountryGermany
Name of the mutant parasite
RMgm numberRMgm-1495
Principal nameĪ”pbanka_1449000
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageNot tested
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of PBANKA_1449000.

Protein (function)
The protein has been identified in an analysis to identify proteins associated with osmiophilic bodies of gametocytes (using a BioID approach of the osmiophilic body protein MDV1/PEG3; see additional information).

Phenotype
Normal numbers of gametocytes and ookinetes are produced. Gametocytes/gametes showed a normal egress phenotype (egress from the host erythrocyte). Ookinetes showed normal motility.

Additional information
This mutant is identified in a study aiming at identifying 'gametocyte regressome', i.e. proteins that are released when Plasmodium gametes escape from their host reythrocyte. For the release of mature extracellular gametes two membrane barriers - the parasite parasitophorous vacuole membrane and the host red blood cell membrane - need to be dissolved. Membrane lysis occurs after the release of proteins from specialized secretory vesicles including osmiophilic bodies.
The identified Plasmodium gametocyte egressome includes the proteins released by the parasite during the lysis of the parasitophorous vacuole membrane and red blood cell membrane. BioID of the osmiophilic body protein MDV1/PEG3 revealed a proteome of these gametocyte-specific secretory vesicles. For bioID the mutated biotin ligase BirA* from Escherichia coli was fused to MDV1/PEG3. BioID relies on the fusion of a protein of interest with a mutated BirA ligase (BirA*) that in the presence of excess biotin biotinylates nearby proteins, which can be purified by affinity chromatography using streptavidin.
Fluorescent protein tagging and gene deletion approaches were employed to validate and identify a set of novel factors essential for this lysis and egress process.

Other mutants
See this link for other mutants generated in this study
RMgm-1488: a mutant expressing a GFP-tagged version of  PBANKA_1449000


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1449000
Gene Model P. falciparum ortholog PF3D7_1234400
Gene productaspartyl protease, putative
Gene product: Alternative name
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 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 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
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