Summary

RMgm-859
Malaria parasiteP. yoelii
Genotype
DisruptedGene model (rodent): PY17X_1210000; Gene model (P.falciparum): PF3D7_1338800; Gene product: CPW-WPC family protein (PyCPW-WPC-1)
PhenotypeNo phenotype has been described
Last modified: 21 April 2013, 20:35
  *RMgm-859
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) : 23587146
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/ResearcherKangwanrangsan, N; Ishino, T.
Name Group/DepartmentDepartment of Molecular Parasitology
Name InstituteGraduate School of Medicine, Ehime University
CityShitsukawa, Toon, Matsuyama, Ehime
CountryJapan
Name of the mutant parasite
RMgm numberRMgm-859
Principal namepycpw-wpc-1(−)
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNot different from wild type
Fertilization and ookineteNot different from wild type
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of PyCPW-WPC-1 (CPW-WPC family protein).

Protein (function)
PyCPW-WPC-1 belongs to a conserved small family (~8 members) named CPW-WPC family (named after the unique WxC motif found at the end of repeated domains). The members share a similar structure, including a signal peptide and five repeated CPW-WPC domains with four to six cysteine residues.

Phenotype
The phenotype analyses indicate that PyCPW-WPC-1 is not essential during the complete life cycle.

Additional information
Evidence is presented for transcription in gametocytes (highly expressed until zygote formation, with a subsequent decrease in expression).
Evidence is presented that PyCPW-WPC-1 protein is produced exclusively in ookinetes. The expression of PyCPW-WPC-1 protein became prominent at 1 and 4 h of ookinete culture and decreased gradually until 24 h of ookinete maturation.
Immunofluorescent staining using mouse anti-PyCPW-WPC-1 antiserum revealed a strong signal on the surface of zygotes, retorts, and mature ookinetes, but not on gametocytes. The staining pattern was similar to that of Pys25, a well-known ookinete surface protein. In addition, another family member, Py03515, also showed a similar surface localization pattern.
PyCPW-WPC-1 is orthologous to P. berghei PBANKA_135250

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PY17X_1210000
Gene Model P. falciparum ortholog PF3D7_1338800
Gene productCPW-WPC family protein
Gene product: Alternative namePyCPW-WPC-1
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid 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 parasitehdhfr
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 15′-CTCGAGTATGCATAATTGTGAATAGTTATTGG-3′
Additional information primer 15'forward
Sequence Primer 25′-GGATCCGACCCCTATAATAATAAAAGGTCTGTC-3′)
Additional information primer 25'reverse
Sequence Primer 35′-AGATCTACCAGATGATTATAATGGACCTTGTCC-3′
Additional information primer 33'forward
Sequence Primer 45′-CTCGAGCGGATGTATTGAGAAGCTTTACATGTG-3′
Additional information primer 43'reverse
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6