RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_0306700; Gene model (P.falciparum): PF3D7_0209600; Gene product: transporter, putative (TRP)
PhenotypeNo phenotype has been described
Last modified: 15 January 2017, 09:23
Successful modificationThe gene/parasite could not be changed/generated by the genetic modification.
The following genetic modifications were attempted Gene disruption
Number of attempts to introduce the genetic modification Unknown
Reference (PubMed-PMID number) Reference 1 (PMID number) : 28027318
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 507cl1 (RMgm-7)
Other information parent lineP.berghei ANKA 507cl1 (RMgm-7) is a reference ANKA mutant line which expresses GFP under control of a constitutive promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190).
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherSrivastava A, Waters AP
Name Group/DepartmentWellcome Centre for Molecular Parasitology, Institute of Infection, Immunity & Inflammation
Name InstituteCollege of Medical, Veterinary and Life Sciences, University of Glasgow

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0306700
Gene Model P. falciparum ortholog PF3D7_0209600
Gene producttransporter, putative
Gene product: Alternative nameTRP
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 modificationThe unsuccessful attempts to knock-out this gene indicate an essential role during blood stage development

From the paper: Viable P.berghei mutants lacking the putative lysine decarboxylase/glutamate decarboxylase (ldc- /gad-, PBANKA_100340o) postulated to synthesise GABA from glutamate were generated and grew at the same rate as wild type asexual parasites, indicating that the limited GABA shunt is not essential for asexual stages. Consistent with this conclusion, viable mutant parasite lines lacking the ornithine amino transferase (OAT), a putative GABA/glutamate transaminase which can recycle glutamate, were also obtained (oat-, PBANKA_0107400) and these parasites grew only slightly less quickly than wild type. However, attempts to generate clonal populations of a mutant (trp-, PBANKA_0306700) lacking a putative GABA transporter (TRP) were unsuccessful, suggesting that this transporter may have a broader role than GABA uptake/efflux. Deletion of another amino transferase (PBANKA_0302300) in P.berghei was previously shown to be refractory
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