RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-5398
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0823800; Gene model (P.falciparum): PF3D7_0922900; Gene product: 3-oxoacyl-[acyl-carrier-protein] reductase (FabG)
Name tag: GFP
Phenotype Oocyst; Sporozoite;
Last modified: 5 March 2024, 18:23
  *RMgm-5398
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
Reference (PubMed-PMID number) Reference 1 (PMID number) : 37543672
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 2.34
Other information parent lineP. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
The mutant parasite was generated by
Name PI/ResearcherSaeed S, Dessens JT
Name Group/DepartmentDepartment of Infection Biology, Faculty of Infectious and Tropical Diseases
Name InstituteLondon School of Hygiene & Tropical Medicine
CityLondon
CountryUK
Name of the mutant parasite
RMgm numberRMgm-5398
Principal nameFabG/GFP
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
OocystNo expression detected in blood stages, gametocytes and ookinetes. FabG expression was readily detected in young oocysts. Fluorescence was localised in a tubular branched structure reminiscent of the apicoplast, in agreement with localisation of FAS in this organelle. FabG::GFP remained apicoplast-localised throughout oocyst development and in sporulated oocysts located in discrete spots, likely refecting the division of the apicoplast tubular network into individual small apicoplasts during sporozoite budding.
SporozoiteFabG::GFP remained apicoplast-localised throughout oocyst development and in sporulated oocysts located in discrete spots, likely refecting the division of the apicoplast tubular network into individual small apicoplasts during sporozoite budding.
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a C-terminal GFP-tagged version of FabG

Protein (function)
FabG is an enzyme of the bacterial like type II fatty acid biosynthesis (FAS-II) pathway

Phenotype
No expression detected in blood stages, gametocytes and ookinetes. FabG expression was readily detected in young oocysts. Fluorescence was localised in a tubular branched structure reminiscent of the apicoplast, in agreement with localisation of FAS in this organelle. FabG::GFP remained apicoplast-localised throughout oocyst development and in sporulated oocysts located in discrete spots, likely refecting the division of the apicoplast tubular network into individual small apicoplasts during sporozoite budding.

Additional information

Other mutants


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0823800
Gene Model P. falciparum ortholog PF3D7_0922900
Gene product3-oxoacyl-[acyl-carrier-protein] reductase
Gene product: Alternative nameFabG
Details of the genetic modification
Name of the tagGFP
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/construct(Linear) plasmid single cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Selectable marker used to select the mutant parasitehdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationTo generate a parasite line expressing FabG fused to GFP (FabG/GFP), an approximately 2.2kb fragment corresponding to the coding sequence (plus introns) and 5’UTR of PBANKA_0823800 was PCR-amplified with primers FabG-F (TTGGGCTGCAGTCGAGGAATTTCCATAGCATCCATATATAC) and FabG-R (AATGAGGGCCCCTAAGCTGGTACCACTTGATAATCCACCATCAATTATAAATAC) and cloned into SalI/HindIII digested pBS-EGFP-hDHFR by In-Fusion to give plasmid pBS-FabG/GFP. This plasmid was linearised with PacI and transfected into purified schizonts for integration into the fabg locus by single crossover homologous recombination
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