RMgmDB - Rodent Malaria genetically modified Parasites


Malaria parasiteP. berghei
MutatedGene model (rodent): PBANKA_144540; Gene model (P.falciparum): PF3D7_1230700; Gene product: protein transport protein SEC13 (SEC13)
Details mutation: The polyproline motif of P. berghei SEC13 replaced by the motif of P. falciparum SEC13
PhenotypeNo phenotype has been described
Last modified: 15 August 2018, 14:52
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene mutation
Reference (PubMed-PMID number) Reference 1 (PMID number) : 30050042
MR4 number
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
The mutant parasite was generated by
Name PI/ResearcherKehrer J, Mair GR, Frischknecht F
Name Group/DepartmentIntegrative Parasitology, Center for Infectious Diseases
Name InstituteHeidelberg University Medical School
Name of the mutant parasite
RMgm numberRMgm-4474
Principal nameSEC13(PF3D7PP)::GFP
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Asexual blood stageNot different from wild type
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

The mutant expresses a C-terminal GFP-tagged version of  a mutated form of SEC13. In P. berghei SEC13 the polyproline motif is replaced by the polyproline motif of P. falciparum SEC13

Protein (function) and Phenotype
In P. berghei the ortholog of SEC13 contains a large C-terminal extension enriched in proline residues. This unordered domain is reminiscent of the C-terminal part in P. berghei SEC31, its binding partner in COPII vesicles. SEC31 also shows a similar, loosely conserved polyproline-rich stretch in P. falciparum and S. cerevisiae. The large, C-terminal extension of SEC13 is conserved in many Plasmodium species and is present in the related apicomplexans.

The number of proline residues in these SEC13 proteins ranges from 73 to 115. While individual proline residues within the SEC13 polyproline stretch of P. berghei are well conserved in the closely related P. yoelii parasite, another rodent malaria species, the amino acid sequence in P. falciparum is highly divergent and aligns poorly.

In order to address a possible function for this C-terminal extension we attempted the generation of a P. berghei mutant lacking this C-terminal polyproline motif. This was not possible indicating that this domain provides an essential role for this protein for parasite survival, either as part of COPII vesicles, or as a Nup, perhaps as an unusual SEC13-Nup145C fusion as suggested for P. falciparum.

Despite the large sequence divergence between P. berghei and P. falciparum, we were readily able to produce a mutant parasite line that expressed the polyproline motif of P. falciparum SEC13 in place of the P. berghei SEC13 polyproline motif. The staining pattern of this protein matched that of wildtype-like P. berghei SEC13-GFP.

Additional information

Other mutants

  Mutated: Mutant parasite with a mutated gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_144540
Gene Model P. falciparum ortholog PF3D7_1230700
Gene productprotein transport protein SEC13
Gene product: Alternative nameSEC13
Details of the genetic modification
Short description of the mutationThe polyproline motif of P. berghei SEC13 replaced by the motif of P. falciparum SEC13
Inducable system usedNo
Short description of the conditional mutagenesisNot available
Additional remarks inducable system
Type of plasmid/construct(Linear) plasmid single cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
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