RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. yoelii
Genetic modification not successful
DisruptedGene model (rodent): PY17X_0203000; Gene model (P.falciparum): Not available; Gene product: integral membrane protein (ETRAMP)
PhenotypeNo phenotype has been described
Last modified: 1 September 2011, 18:19
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 ≥ 5
Reference (PubMed-PMID number) Reference 1 (PMID number) : 21819513
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. yoelii
Parent strain/lineP. y. yoelii 17XNL
Name parent line/clone Not applicable
Other information parent line17XNL is a non-lethal strain of P. yoelii
Attempts to generate the mutant parasite were performed by
Name PI/ResearcherD.C. MacKellar; S.H.I. Kappe
Name Group/DepartmentMolecular and Cellular Biology Program
Name InstituteUniversity of Washington
CitySeattle, Washington

  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PY17X_0203000
Gene Model P. falciparum ortholog Not available
Gene productintegral membrane protein
Gene product: Alternative nameETRAMP
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, SpeI
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 modificationThe unsuccessful attempts to disrupt the gene indicates that PY02667 is essential for blood stage growth/multiplication.

The replacement of PY02667 with an epitope-tagged copy (RMgm-640) suggests this failure was not due to inaccessibility of the genomic locus.

Early transcribed membrane protein (ETRAMP) family member. Plasmodium conserved family with greater than ten members in P. falciparum. ETRAMPs are abundantly expressed early in the intraerythrocytic cycle and are small (frequently less than 200 aa) integral membrane proteins that are localized within the parasitophorous vacuolar membrane (PVM). All members have signal peptides plus a transmembrane domain. The ETRAMP/SEP proteins of P. yoelii and P. berghei (8-11 genes) show homology to members of the ETRAMP family of proteins of P. falciparum (14 genes) but the orthologous relationship of the different members is not completely resolved.
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
Additional information primer 12667 Rep1F (5')
Additional information primer 22667 Rep2R (5')
Additional information primer 32667 Rep3F (3')
Additional information primer 42667 Rep4R
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6