RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_0833000; Gene model (P.falciparum): PF3D7_0932200; Gene product: profilin (Pfn)
PhenotypeNo phenotype has been described
Last modified: 18 February 2010, 18:56
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 3
Reference (PubMed-PMID number) Reference 1 (PMID number) : 19000816
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255)
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherI. Kursula; H. Schüler
Name Group/DepartmentDepartment of Biochemistry
Name InstituteUniversity of Oulu

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0833000
Gene Model P. falciparum ortholog PF3D7_0932200
Gene productprofilin
Gene product: Alternative namePfn
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 SacII, KpnI
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 modificationNo detailed information on primer sequences is provided.

Attempts to disrupt the profilin gene of P. berghei were unsuccessful suggesting an essential role during blood stage development.

Profilins are ubiquitous and essential actin monomer binding proteins, known to interact with proline-rich regions in a variety of proteins and to regulate actin filament formation. For fast actin polymerization at selected sites, profilin is an essential control element by recruiting actin monomers in a polymerizable form to actin polymerization machineries. Profilins are usually defined by their shared, highly conserved structure and by their ability to bind actin, proline-rich sequences

The genome of the different Plasmodium spcies contains a 'profilin' ortholog

In this study it is shown that profilin of P. falciparum adopts an overall core structure and displays biochemical properties that together allow for its classification as a conventional profilin. However, it also possesses a unique minidomain, including an arm-like b-hairpin protrusion common to apicomplexan parasites, and an acidic loop specific for Plasmodium species. Profilin is an essential gene for Plasmodium blood stages. In this study a control replacement plasmid was generated that contained a functional full-length copy of the human profilin gene (Pfn1) under the control of the endogenous PbPfn promoter and a constitutive untranslated region to permit proper transcript stability. Transfection of this construct yielded viable parasites that contained the predicted allelic replacement at the first attempt. These findings show that (i) the PbPfn locus is readily accessible to homologous recombination, and (ii) human profilin can complement the essential role of the parasite protein.
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