Summary

RMgm-1327
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0931200; Gene model (P.falciparum): PF3D7_1116800; Gene product: heat shock protein 101|chaperone protein ClpB2 (HSP101)
Name tag: mCherry-3xMyc
Phenotype Asexual bloodstage;
Last modified: 5 April 2016, 09:33
  *RMgm-1327
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
Reference (PubMed-PMID number) Reference 1 (PMID number) : 26219962
MR4 number
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).
The mutant parasite was generated by
Name PI/ResearcherMatz, JM; Kooij, TW
Name Group/DepartmentMedical Microbiology
Name InstituteRadboudumc
CityNijmegen
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-1327
Principal namehsp101-mCh(sens)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageAbundant expression at the PVM in younger blood stages and in motile tubular structures in maturing trophozoites
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant expresses a mCherry tagged version of HSP101 (and lacks the hdfr-yfcu drug-selectable marker that has been removed by negative selection)

Protein (function)
Plasmodium parasites remodel their vertebrate host cells by translocating hundreds of proteins across an encasing membrane into the host cell cytosol via a putative export machinery termed PTEX (Plasmodium Translocon of EXported protein). HSP101 (PbANKA_094120), PTEX150 (PbANKA_100850), EXP2 (PbANKA_133430), PTEX88 (PbANKA_094130) and TRX2 (PbANKA_135800) have been identified as members of the PTEX complex.

Phenotype
Phenotype analyses indicate that HSP101 is located at the PVM of blood stages.

Additional information
In the paper a mutant is described that expresses mCherry-tagged HSP101 and expresses GFP in the parasitophorous vacuole (PV). This mutant is obtained by (mosquito) crossing of mutant RMgm-1327 with mutant RMgm-1333

In the paper a mutant is described that expresses mCherry-tagged HSP101 and expresses GFP in the endoplasmatic reticulum (ER). This mutant is obtained by (mosquito) crossing of mutant RMgm-1327 with mutant RMgm-1332

Other mutants
See link for other HSP101 mutants


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0931200
Gene Model P. falciparum ortholog PF3D7_1116800
Gene productheat shock protein 101|chaperone protein ClpB2
Gene product: Alternative nameHSP101
Details of the genetic modification
Name of the tagmCherry-3xMyc
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodies
Type of plasmid/construct(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
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedure5-fluorocytosine (5-FC)
Additional remarks genetic modificationThe hdfr-yfcu drug-selectable marker has been removed by negative selection with 5-FC
Additional remarks selection procedureThe hdfr-yfcu drug-selectable marker has been removed by negative selection with 5-FC
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