RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-926
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0501400; Gene model (P.falciparum): PF3D7_1017100; Gene product: rhoptry neck protein 12 (RON12)
Phenotype Asexual bloodstage;
Last modified: 17 August 2013, 15:05
  *RMgm-926
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) : 23937520
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/ResearcherKnuepfer, E; Tewari, R.; Holder, A.A.
Name Group/DepartmentDivision of Parasitology
Name InstituteMRC National Institute for Medical Research, Mill Hill
CityLondon
CountryUK
Name of the mutant parasite
RMgm numberRMgm-926
Principal namePbΔron12
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageParasitemia developed more slowly in mice infected with PbΔron12 compared to wild-type PbANKA showing a delay of approximately one day before reaching similar parasitemias as the wild type with the curves otherwise showing a highly similar profile.
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of RON12

Protein (function)
RON12 is found only in Plasmodium and is highly conserved across the genus. RON12 lacks membrane anchors and is a major soluble component of the nascent PV. The bulk of the secretion of RON12 happens late during invasion (after parasite internalisation) allowing accumulation in the fully formed PV with a small proportion of RON12 also apparent in structures resembling the moving junction.
Gene deletion mutants show (slightly) reduced asexual blood stage proliferation in both P. falciparum and in P. berghei.

Phenotype
Parasitemia developed more slowly in mice infected with PbΔron12 compared to wild-type PbANKA showing a delay of approximately one day before reaching similar parasitemias as the wild type with the curves otherwise showing a highly similar profile.

Additional information

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0501400
Gene Model P. falciparum ortholog PF3D7_1017100
Gene productrhoptry neck protein 12
Gene product: Alternative nameRON12
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 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 15’-AGTTAAGGTACCCGCATTGCCTAGACATTGG-3’
Additional information primer 1PbRON12F1for (5')
Sequence Primer 25’- GCTGCAAGCTTCTTTGTATATTTCCCTATCC-3’
Additional information primer 2PbRON12F1rev (5')
Sequence Primer 35’-AGCCGCGGAATTCGACCGAATAAAAGTACCTAATTGC-3
Additional information primer 3PbRON12F2for (3')
Sequence Primer 45’-CTAATTGCGGCCGCCACATGCTACTAACGTCGCC-3’
Additional information primer 4PbRON12F2rev (3')
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6