RMgmDB - Rodent Malaria genetically modified Parasites

Back to search results

Summary

RMgm-1593
Malaria parasiteP. berghei
Genotype
Transgene
 Transgene Plasmodium: Gene model: PBANKA_0904100; Gene model (P.falciparum): PF3D7_1144900; Gene product: ras-related protein Rab-6 (RAB6)
Promoter: Gene model: PBANKA_1003000; Gene model (P.falciparum): PF3D7_0405300; Gene product: liver specific protein 2, putative | sequestrin | 6-cysteine protein (LISP2)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Insertion locus: Gene model: Not available; Gene product: Not available (small subunit ribosomal rna gene (c/d-type unit))
Phenotype Liver stage;
Last modified: 21 September 2016, 15:10
  *RMgm-1593
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 27566438
MR4 number
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
The mutant parasite was generated by
Name PI/ResearcherKaiser G; Heussler VT; Stanway RR
Name Group/DepartmentInstitute of Cell Biology
Name InstituteUniversity of Bern
CityBern
CountrySwitzerland
top of page
Name of the mutant parasite
RMgm numberRMgm-1593
Principal namePblsGFP-Rab6
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
top of page
Phenotype
Asexual blood stageNot tested
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageFor analysis of the trans-Golgi localisation, PbLSGFP-Rab6 parasites were stained with ER-TrackerTM Red and imaged live. It was found that trans-Golgi faces are localised at ER accumulations but are also frequently located at other regions of the ER.
Additional remarks phenotype

Mutant/mutation
The mutant expresses a GFP-tagged version of P. berghei Rab6. The transgene is integrated into the silent c/d-type rrna gene unit under control of the liver stage specific promoter lisp2.

Protein (function)
In the cells of most organisms, the cis-Golgi is found juxtaposed to the tER, to allow effective COPII-mediated protein transport between the ER and Golgi apparatus. As Plasmodium parasites possess a non-classical unstacked Golgi, the trans-face of the Golgi is found in close proximity to the cis-Golgi. The Golgi reassembling stacking protein (GRASP) and Rab6 are well-established marker proteins of the cis-Golgi and trans-Golgi, respectively, and have been used in P. falciparum for visualisation of both Golgi faces in the parasite’s blood stage.

Phenotype
For analysis of the trans-Golgi localisation, PbLSGFP-Rab6 parasites were stained with ER-TrackerTM Red and imaged live. It was found that trans-Golgi faces are  localised at ER accumulations but are also frequently located at other regions of the ER.

See also  RMgm-1588 for a mutant expresses an N-terminal GFP-tagged version of P. berghei Sec61β. Sec61β is a known marker protein for endoplasmic reticulum (ER). 

Additional information

Other mutants

  Transgene: Mutant parasite expressing a transgene
top of page
Type and details of transgene
Is the transgene Plasmodium derived Transgene: Plasmodium
Gene Model of Parasite PBANKA_0904100
Gene Model P. falciparum ortholog PF3D7_1144900
Gene productras-related protein Rab-6
Gene product: Alternative nameRAB6
top of page
Details of the genetic modification
Inducable system usedNo
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 modificationcDNA of of PbRab6 (PBANKA_090410) was amplified using the primers 5’- GTA CTC TAG AAT GGA CGA ATT TCA AAA TTC TG-3’ and 5’- TCG ATC TAG ATT AAC ACA AAC ATT TAC TTA GC-3’. The PbRab6 fragment was inserted using XbaI into a modified version of pGFP103464 (Helm et al., 2010), containing a GFP without stop codon to facilitate N-terminal cloning.
Additional remarks selection procedure
top of page
Other details transgene
top of page
Promoter
Gene Model of Parasite PBANKA_1003000
Gene Model P. falciparum ortholog PF3D7_0405300
Gene productliver specific protein 2, putative | sequestrin | 6-cysteine protein
Gene product: Alternative nameLISP2
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
top of page
3'-UTR
Gene Model of Parasite PBANKA_0719300
Gene productbifunctional dihydrofolate reductase-thymidylate synthase, putative
Gene product: Alternative namedhfr/ts
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Insertion/Replacement locus
Replacement / InsertionInsertion locus
Gene Model of Parasite Not available
Gene productNot available
Gene product: Alternative namesmall subunit ribosomal rna gene (c/d-type unit)
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the 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
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