RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-4986
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0404100; Gene model (P.falciparum): PF3D7_0305600; Gene product: DNA-(apurinic or apyrimidinic site) endonuclease (APE1)
Phenotype Liver stage;
Last modified: 13 May 2021, 17:25
  *RMgm-4986
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) : 33743509
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/ResearcherVerma N, Habib S
Name Group/DepartmentDivision of Molecular and Structural Biology
Name InstituteCSIR-Central Drug Research Institute
CityLucknow
CountryIndia
Name of the mutant parasite
RMgm numberRMgm-4986
Principal nameAPE1(-)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
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 stageProlonged prepatent period after infecting mice intravenously with sporozoites (prolonged with 2 days).
In cultured heaptocytes Ape1(-) parasites develop into EEFs and egress from hepatocytes normally. Normal merosome formation and segregation of merozoites. Evidence is presented the merozoites from merosomes have decreased infectivity.
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of APE1 and expresses GFP

Protein (function)
The protein is an ExoIII family endonucleases (proteins involved in organellar base excision repair (BER pathway). The P. falciparum nuclear genome encodes two class II AP  endonucleases– an endonuclease IV (EndoIV/Nfo) family protein (PfApn1) and an exonuclease III (ExoIII) homolog (PfApe1) The parasite ExoIII family endonuclease, PfApe1,  has a strong mitochondrial targeting prediction.

Phenotype
Prolonged prepatent period after infecting mice intravenously with sporozoites (prolonged with 2 days).
In cultured heaptocytes Ape1(-) parasites develop into EEFs and egress from hepatocytes normally. Normal merosome formation and segregation of merozoites. Evidence is presented the merozoites from merosomes have decreased infectivity.

Additional information
Evidence is presented that: PfApe1 localized in the mitochondrion and exhibited AP-site cleavage, 3′-5′ exonuclease, 3′-phosphatase, nucleotide incision repair (NIR) and RNA cleavage activities

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0404100
Gene Model P. falciparum ortholog PF3D7_0305600
Gene productDNA-(apurinic or apyrimidinic site) endonuclease
Gene product: Alternative nameAPE1
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 parasitehdhfr/yfcu
Promoter of the selectable markereef1a
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