SummaryRMgm-587
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Successful modification | The 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) : 20852334 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei ANKA |
Name parent line/clone | P. berghei ANKA cl15cy1 |
Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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The mutant parasite was generated by | |
Name PI/Researcher | K. Buchholz; K. Becker; K. Matuschewski |
Name Group/Department | Interdisciplinary Research Centre |
Name Institute | Justus-Liebig University |
City | Giessen |
Country | Germany |
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Name of the mutant parasite | |
RMgm number | RMgm-587 |
Principal name | GR(-) |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | The mutant showed a slight delay in initial proliferation of blood stages in mice. |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Not different from wild type |
Oocyst | Mutants produced lower numbers of oocysts in comparison with wild type parasites (although the differences were not significant). Mutant oocysts were significantly smaller than wild type oocysts and no sporozoites were formed. |
Sporozoite | No production of oocyst- or salivary gland sporozoites |
Liver stage | No production of oocyst- or salivary gland sporozoites. Infected mosquitoes were unable to infect mice by mosquito bite |
Additional remarks phenotype | Mutant/mutation Through disruption of the gene encoding γ-GCS it has been shown that de novo GSH synthesis is not critical for P. berghei blood stage multiplication but is essential for oocyst development. The phenotype analyses of mutant parasites lacking expression of GR confirms that GSH metabolism is critical for the mosquito oocyst stage. See also the P. berghei mutants RMgm-403 and RMgm-404 which also lack the expression of GR and which show a similar phenotype as the mutant described here. |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1023400 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1419800 | ||||||||||||||||||||||||
Gene product | glutathione reductase | ||||||||||||||||||||||||
Gene product: Alternative name | GR | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | Plasmid double cross-over | ||||||||||||||||||||||||
PlasmoGEM (Sanger) construct/vector used | No | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||
Restriction sites to linearize plasmid | SacII, KpnI | ||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | tgdhfr | ||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
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