Summary

RMgm-4459
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_0926200; Gene model (P.falciparum): PF3D7_1122100; Gene product: GPI transamidase component GPI16, putative (GPI16)
Phenotype Asexual bloodstage;
Last modified: 29 June 2018, 18:22
  *RMgm-4459
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) : 29784863
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/ResearcherLiu Q; Cao Y
Name Group/DepartmentDepartment of Immunology, College of Basic Medical Sciences
Name InstituteChina Medical University Shenyang
CityShenyang, Liaoning
CountryChina
Name of the mutant parasite
RMgm numberRMgm-4459
Principal nameΔpbgpi16
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageC57Bl6 mice were infected with Δpbgpi16 and wild type (WT) PbA. In both groups parasitemia rose steadily after infection and did not show statistically significant difference by day 7 (P > 0.05, two-way ANOVA), albeit the day 7 parasitemia in the WT-infected group (7.4%) was higher than that in the Δpbgpi6-infected mice (6.0%).
Reduced mortality due to complications of ECM in Δpbgpi16 infected mice.
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of GPI16

Protein (function)
Glycosylphosphatidylinositols (GPIs) and GPI anchors are transported to the cell surface via a process called GPI transamidation, which involves the GPI transamidase (GPI-T) complex. Before GPIs and GPI anchors are transported to the cell surface, the GPI attachment signal peptide is replaced by a preassembled GPI in the endoplasmic reticulum through a transamidation reaction that results in the formation of a carbonyl intermediate. The GPI transamidase is a key enzyme of this posttranslational modification. The GPI transamidase complex consists of five different subunits and is conserved among different species. In this study, we characterized the GPI transamidase subunit16 (Gpi16)  (PBANKA_0926200) homolog in P. berghei (PbGPI16), a highly conserved GPI-T subunit among Plasmodium species.

Phenotype
C57Bl6 mice were infected with Δpbgpi16 and wild type (WT) PbA. In both groups parasitemia rose steadily after infection and did not show statistically significant difference by day 7 (P > 0.05, two-way ANOVA), albeit the day 7 parasitemia in the WT-infected group (7.4%) was higher than that in the Δpbgpi6-infected mice (6.0%).
Reduced mortality due to complications of ECM in Δpbgpi16 infected mice.

Additional information
Evidence is presented that:
- total GPIs in blood stages of Δpbgpi16 were slightly less than in wild type (WT) PbA
- GPIs associated with proteins extracted from merozoite membranes were significantly less in Δpbgpi16 than in WT parasites
- Proteins were analyzed in Western blots using antibodies against PbMSP1, PbMSP4 and PbMSP10, respectively. The results showed that the amounts of all three proteins were significantly reduced in the merozoite membrane of Δpbgpi16 parasites.

Other mutants


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0926200
Gene Model P. falciparum ortholog PF3D7_1122100
Gene productGPI transamidase component GPI16, putative
Gene product: Alternative nameGPI16
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 parasitehdhfr
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