Summary

RMgm-4834
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0608000; Gene model (P.falciparum): PF3D7_1209600; Gene product: porphobilinogen deaminase (PBGD)
PhenotypeNo phenotype has been described
Last modified: 22 June 2020, 12:56
  *RMgm-4834
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Not published (yet)
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/ResearcherSchnider CB, McMorran BJ
Name Group/DepartmentDepartment of Immunology and Infectious Disease, John Curtin School of Medical Research
Name InstituteThe Australian National University
CityCanberra
CountryAustralia.
Name of the mutant parasite
RMgm numberRMgm-4834
Principal namePbA_pbgd
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of PBGD

Protein (function)
The malaria parasite is capable of de novo heme biosynthesis despite its ability to acquire heme from red blood cell (RBC) hemoglobin. The first enzyme of the de novo heme-biosynthetic pathway, δ-aminolevulinate synthase (ALAS) and the last two enzymes, Protoporphyrinogen IX oxidase (PPO) and Ferrochelatase (FC) localize to the mitochondrion. The enzymes that catalyze the intermediate steps: ALA dehydratase (ALAD), Porphobilinogen deaminase (PBGD) and Uroporphyrinogen III decarboxylase (UROD) localize to the apicoplast.

Phenotype
Normal asexual blood stage growth/multiplication in mice

Additional information
From the Abstract (published in bioRxiv preprint doi: https://doi.org/10.1101/589242. This version posted June 13, 2020):
'A mouse ENU-induced mutagenesis screen for novel malaria resistance-conferring mutations identified a novel nonsense mutation in the gene encoding porphobilinogen deaminase (PBGD) in mice, denoted here as PbgdMRI58155. Heterozygote PbgdMRI58155 mice exhibited approximately 50% reduction in cellular PBGD activity in both mature erythrocytes and reticulocytes, although enzyme activity was approximately 10 times higher in reticulocytes than erythrocytes. When challenged with blood-stage P. chabaudi, which preferentially infects erythrocytes, heterozygote mice showed a modest but significant resistance to infection, including reduced parasite growth.
The Plasmodium resistance phenotype was not recapitulated in Pbgd-deficient mice infected with P. berghei, which prefers reticulocytes, or when P. falciparum was cultured in erythrocytes from patients with acute intermittent porphyria (AIP), which had modest (20-50%) reduced levels of PBGD. Furthermore, the growth of Pbgd-null P. falciparum and Pbgd-null P. berghei parasites, which grew at the same rate as their wild-type counterparts in normal cells, were not affected by the PBGD-deficient background of the AIP erythrocytes or Pbgd-deficient mice'.

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0608000
Gene Model P. falciparum ortholog PF3D7_1209600
Gene productporphobilinogen deaminase
Gene product: Alternative namePBGD
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