RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-4456
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0603300; Gene model (P.falciparum): PF3D7_1204400; Gene product: conserved Plasmodium membrane protein, unknown function (Pbg37)
Phenotype Gametocyte/Gamete; Fertilization and ookinete; Oocyst;
Last modified: 6 June 2018, 17:55
  *RMgm-4456
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) : 29866905
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/ResearcherLiu F, Cao Y
Name Group/DepartmentDepartment of Immunology, College of Basic Medical Sciences
Name InstituteChina Medical University
CityShenyang, Liaoning
CountryChina
Name of the mutant parasite
RMgm numberRMgm-4456
Principal nameΔpbg37
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteReduced gametocyte production. A higher female/male gametocyte ratio, fewer mature male gametocytes and defects in exflagellation of mature microgametocytes.
Fertilization and ookinetereduced (75%) ookinete formation
OocystMosquitoes fed on mice infected with the Δpbg37 lines showed a modest reduction in infection rate (68.2% in the Δpbg37 groups) compared to those fed on mice infected with the WT parasites (93.1%). Moreover, there was a 91.4% reduction in the mean oocyst number/midgut in mosquitoes fed on mice infected with the Δpbg37 parasites (12.2) as compared to that in the WT group (140.9).
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of Pbg37

Protein (function)
A protein of 37 kDa with a signal peptide and 34 multiple transmembrane domains, and preferentially expressed in gametocytes.

Phenotype
Reduced gametocyte production and reduced ookinete and oocyst formation.

Additional information
Recombinant protein (rPbg37) targeting the N-terminal 63 amino acids (26-88) expressed in bacteria elicited strong antibody responses in mice. Western blots demonstrated Pbg37 expression in gametocytes, zygotes, and to a lesser extent, ookinetes, and its predominant association with membranes of gametocytes. Indirect immunofluorescence assay showed abundant surface localization of Pbg37 on gametes and zygotes, but reduced amounts on retorts and ookinetes.

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0603300
Gene Model P. falciparum ortholog PF3D7_1204400
Gene productconserved Plasmodium membrane protein, unknown function
Gene product: Alternative namePbg37
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) PCR construct 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 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