RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-971
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1003900; Gene model (P.falciparum): PF3D7_0406200; Gene product: sexual stage-specific protein precursor (Pfs16)
Name tag: triple-HA
PhenotypeNo phenotype has been described
Last modified: 14 December 2018, 11:11
  *RMgm-971
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
Reference (PubMed-PMID number) Not published (yet)
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 2.34
Other information parent lineP. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943).
The mutant parasite was generated by
Name PI/ResearcherSiden-Kiamos, I.
Name Group/DepartmentInstitute of Molecular Biology and Biotechnology
Name InstituteFORTH
CityHeraklion
CountryGreece
Name of the mutant parasite
RMgm numberRMgm-971
Principal name100390-HA
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
In the mutant the PBANKA_1003900 gene has been (C-terminally) tagged with a triple-HA tag using a PlasoGem DNA-construct.
This gene is a 'syntenic ortholog' of sexual stage-specific protein precursor (Pfs16; PF3D7_0406200) of P. falciparum.

The phenotype has not been analysed in detail. No tagged-protein could be detected by immuno-fluorescence analyses in asexual blood stages, gametocytes and ookinetes.

An additional mutant has been generated that contains the PBANKA_1003900 gene (C-terminally) tagged with GFP on episomes. The tagged-gene is under the control of ~1kb 5'UTR  region of PBANKA_1003900 and the 3'UTR of the pbdhfr/ts gene. Also in this mutant no tagged-protein could be detected in asexual blood stages, gametocytes and ookinetes.

The similarity of P. falciparum Pfs16 and the syntenic P. berghei gene (Pbs16?) is below threshold.

See also RMgm-968 for a mutant with a disrupted PBANKA_100390) gene locus
See the link PBANKA_1003900 for other mutants

 


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1003900
Gene Model P. falciparum ortholog PF3D7_0406200
Gene productsexual stage-specific protein precursor
Gene product: Alternative namePfs16
Details of the genetic modification
Name of the tagtriple-HA
Details of taggingC-terminal
Additional remarks: taggingThe PBANKA_100390 gene has been tagged using a PlasoGem DNA-construct
See the PlasmoGem website for details of the DNA construct
Commercial source of tag-antibodies
Type of plasmid/construct(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedYes
Name of PlasmoGEM construct/vector95547
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 markereef1a
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