Summary

RMgm-4576
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1224400; Gene model (P.falciparum): PF3D7_0806300; Gene product: ferlin, putative (ferlin-like protein, FLP)
PhenotypeNo phenotype has been described
Last modified: 2 January 2019, 15:34
  *RMgm-4576
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) : 30597708
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/ResearcherObrova K, Mair GR, Mueller AK
Name Group/DepartmentCenter for Infectious Diseases, Parasitology Unit
Name InstituteHeidelberg University Hospital
CityHeidelberg
CountryGermany

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1224400
Gene Model P. falciparum ortholog PF3D7_0806300
Gene productferlin, putative
Gene product: Alternative nameferlin-like protein, FLP
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 parasitetgdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationFrom the paper;

'Attempts to delete flp using standard gene targeting approaches with the plasmid b3D.DT∧H.∧D or PlasmoGEM vectors however failed repeatedly in our hands, and a recent global screen identified the gene to be essential for the blood stage. This strongly suggests that FLP fulfils an essential function during the intra-erythrocytic development cycle.

The P. berghei ferlin-like protein (FLP, PBANKA_1224400) contains six predicted C2 domains, a C-terminal transmembrane domain and a Fer domain (a ferlin-specific motif of unknown function). These typical features cluster FLP together with ferlin (PBANKA_1319300) into the ferlin protein family.Ferlins are typically embedded in vesicular membranes via a single, C-terminal transmembrane domain with the N-terminus of the protein facing the cytosol.'
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