Summary

RMgm-4444
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_1213100; Gene model (P.falciparum): PF3D7_1014700; Gene product: prohibitin, putative (PHB2)
PhenotypeNo phenotype has been described
Last modified: 27 April 2018, 17:41
  *RMgm-4444
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) : 29669282
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
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherMatz JM, Kooij TWA
Name Group/DepartmentDepartment of Molecular Parasitology, Institute of Biology
Name InstituteHumboldt University
CityBerlin
CountryGermany

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1213100
Gene Model P. falciparum ortholog PF3D7_1014700
Gene productprohibitin, putative
Gene product: Alternative namePHB2
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 markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modificationThe unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood stage growth

Proteins of the stomatin/prohibitin/flotillin/HfIK/C (SPFH) family are membrane-anchored and perform diverse cellular functions in different organelles. A systematic database search for Plasmodium SPFH proteins uncovered four candidates that either display a close phylogenetic relationship to conserved members—prohibitin 1 (PHB1, PBANKA_0701500), prohibitin 2 (PHB2, PBANKA_1213100), and stomatin-like protein (STOML, PBANKA_0807100)- 0807100)—or fall into a unique sister group that contains related proteins from the phylum Myzozoa, which harbors the subphylum Apicomplexa. Accordingly, the latter Plasmodium protein was named prohibitin-like protein (PHBL, PBANKA_0718600). All four proteins contain a central signature SPFH domain and a hydrophobic stretch Although the conventional Plasmodium SPFH proteins display a similarly high degree of conservation, PHBL sequences are rather divergent already among related Plasmodium species.
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