Summary

RMgm-4410
Malaria parasiteP. yoelii
Genotype
DisruptedGene model (rodent): PY17X_1451500; Gene model (P.falciparum): PF3D7_1234400; Gene product: microgamete surface protein MiGS, putative (MiGS; aspartyl protease, putative)
Phenotype Gametocyte/Gamete; Fertilization and ookinete; Oocyst;
Last modified: 11 January 2018, 17:44
  *RMgm-4410
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 29316140
MR4 number
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 line
The mutant parasite was generated by
Name PI/ResearcherTachibana M, Torii M
Name Group/DepartmentDivision of Molecular Parasitology, Proteo-Science Center
Name InstituteEhime University
CityToon Ehime
CountryJapan
Name of the mutant parasite
RMgm numberRMgm-4410
Principal nameΔPyMiGS
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNormal gametocyte production. Reduced osmiophilic body (OB) formation in male gametocytes. Normal gametocyte egress from the erythrocyte. Reduced numbers of exflagellating males after RBC egress.
Fertilization and ookineteReduced fertilisation/ookinete formation.
OocystReduced fertilisation/ookinete formation. Reduced oocyst formation.
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of MiGS

Protein (function)
PyMiGS (PY17X_1451500) is a 594 amino acid protein containing an N-terminal signal peptide, aspartyl protease-like domain, and a C-terminal transmembrane domain. The aspartyl protease-like domain is conserved among MiGS orthologues in other Plasmodium species.

Phenotype
Normal gametocyte production. Reduced osmiophilic body (OB) formation in male gametocytes. Normal gametocyte egress from the erythrocyte. Reduced numbers of exflagellating males after RBC egress. Reduced fertilisation/ookinete formation.

Additional information
Evidence is presented:
- that MiGS is expressed in male gametocytes and microgametes (and not in female gametoctyes).
- PyMiGS is localized to the osmiophilic body (OB) and microgamete surface
- PyMiGS is important for OB formation in male gametocytes (reduced OB formation in the knock-out mutant)
- PyMiGS is dispensable for gametocyte egress from the erythrocyte.
- PyMiGS plays an important role in exflagellation. 66% of PyWT male activated gametocytes performed exflagellation, whereas 14% to 35% of ΔPyMiGS male activated gametocytes exflagellated
- In the case of PyWT, 91% of female gametocytes transformed to zygotes or further developed to ookinetes. In contrast, only 34% of ΔPyMiGS clone #1 and 57% of ΔPyMiGS clone #2 transformed to zygote or ookinetes
- The oocyst numbers of ΔPyMiGS on day 10 post-feeding were also decreased compared with those of PyWT. This reduction was comparable to the reduction of zygote/ookinete conversion rate observed in the ΔPyMiGS gametes.

Other mutants
See the link PF3D7_1234400 for other mutants (with slight differences in phenotypes)


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PY17X_1451500
Gene Model P. falciparum ortholog PF3D7_1234400
Gene productmicrogamete surface protein MiGS, putative
Gene product: Alternative nameMiGS; aspartyl protease, putative
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(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
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 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