Summary

RMgm-4832
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0414500; Gene model (P.falciparum): PF3D7_0316700; Gene product: protein YOP1, putative (YOP1)
Phenotype Asexual bloodstage; Oocyst; Sporozoite; Liver stage;
Last modified: 21 June 2020, 21:22
  *RMgm-4832
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) : 32432369
MR4 number
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
The mutant parasite was generated by
Name PI/ResearcherShi X, Bhanot P
Name Group/DepartmentDepartment of Microbiology, Biochemistry and Molecular Genetics
Name InstituteRutgers New Jersey Medical School
CityNewark, NJ
CountryUSA
Name of the mutant parasite
RMgm numberRMgm-4832
Principal namePbyop1∆
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageReduced growth of asexual blood stages. Pbyop1∆ parasites have a multiplication rate of approximately 7-fold over 24 h compared to 10-fold for 127 WT parasites.
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystReduced oocysts and sporozoite loads. Partially (?) explained by reduced growth of blood stages.
SporozoiteReduced oocysts and sporozoite loads. Partially (?) explained by reduced growth of blood stages.
Liver stage(Some) Evidence for reduced liver stage development
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of YOP1

Protein (function)
Rodent-infective and human-infective Plasmodium species encodes three homologs of proteins that induce membrane curvature and form ER tubules in higher eukaryotes, YOP1 (PF3D7_0316700 ), YOP1-like (YOP1L; PF3D7_1358700) and RTN1 (PBANKA_1139900). The primary structures of YOP1, YOP1L and RTN1 are highly conserved amongst rodent and human Plasmodium species (71-73% identity in amino acid sequence).

Phenotype
Reduced growth of asexual blood stages. Pbyop1∆ parasites have a multiplication rate of approximately 7-fold over 24 h compared to 10-fold for 127 WT parasites.
Reduced oocysts and sporozoite loads. Partially (?) explained by reduced growth of blood stages.
(Some) Evidence for reduced liver stage development.

Evidence is presented that:
- YOP1 proteins may play an evolutionarily conserved role in maintaining ER and vacuole morphology

Additional information
IFA analyses showed: In asexual stages, PbYOP1 is present in subcellular structures consistent with the peripheral ER. As observed with yeast and mammalian homologs, PbYOP1 partially co-localizes with a pan-ER marker BiP1, a protein resident in the ER lumen. In sporozoites, PbYOP1’s localization is primarily perinuclear, consistent with ER localization. PbYOP1 was not detectable in liver stages 48 h post infection

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0414500
Gene Model P. falciparum ortholog PF3D7_0316700
Gene productprotein YOP1, putative
Gene product: Alternative nameYOP1
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 usedYes
Name of PlasmoGEM construct/vectorpBGEM-272858
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/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