RMgmDB - Rodent Malaria genetically modified Parasites

Back to search results

Summary

RMgm-1587
Malaria parasiteP. berghei
Genotype
Genetic modification not successful
DisruptedGene model (rodent): PBANKA_0105900; Gene model (P.falciparum): PF3D7_1440300; Gene product: delta-aminolevulinic acid dehydratase | porphobilinogen synthase (PBGS; ALAD)
PhenotypeNo phenotype has been described
Last modified: 12 September 2016, 19:13
  *RMgm-1587
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) : 27600503
top of page
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
top of page
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherRizopoulos Z, Matuschewski K, Haussig J.
Name Group/DepartmentParasitology Unit
Name InstituteMax Planck Institute for Infection Biology
CityBerlin
CountryGermany
  Disrupted: Mutant parasite with a disrupted gene
top of page
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0105900
Gene Model P. falciparum ortholog PF3D7_1440300
Gene productdelta-aminolevulinic acid dehydratase | porphobilinogen synthase
Gene product: Alternative namePBGS; ALAD
top of page
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 modificationUnsuccessful attempts to disrupt the gene indicate that this gene is essential for asexual blood stage growth/multiplication.

Fragments of the 3’ and 5’ UTR were amplified from gDNA using gene-specific primers. First, 3’ fragments were cloned into the pBAT vector, following restriction digestion with HindIII and KpnI to generate an intermediate construct (pHEME-im). Then, the 5’ UTR fragment was digested with SacII and EcoRV and cloned into the respective SacII- and PvuII-linearized pHEME-im construct, thus removing the mCherry-3xMyc tag. Finally, the pHEME-ko vector was linearized with ScaI and SalI before transfection into P. berghei strain ANKA (wt) parasites.
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
top of page
Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: StudioDrie StudioDrie; S. Mededovic and A. van Wigcheren