SummaryRMgm-46
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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) : 16468982 |
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 | Not applicable |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | T. Kariu, M. Yuda |
Name Group/Department | School of Medicine |
Name Institute | Mie University |
City | Mie |
Country | Japan |
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Name of the mutant parasite | |
RMgm number | RMgm-46 |
Principal name | celtos(-)1; celtos(-)2; celtos(-)3 |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | Not different from wild type |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Normal production of gametocytes. Normal fertilization rate and ookinete production. ookinete infectivity is decreased (200 fold reduction in oocyst production). Ookinetes penetrate the epithelial cells of the mosquito midgut but are affected in migration through the cytoplasm of the epithelial cell to the basal lamina. |
Oocyst | The number of oocysts is decreased by ~200 fold compared to wild type as a result of decreased infectivity of ookinetes (see phenotype 'Fertilization and ookinete'). Oocysts produce normal numbers of sporozoites that are not affected in their infectivity to salivary glands. |
Sporozoite | The number of oocysts is decreased by ~200 fold compared to wildtype as a result of decreased infectivity of ookinetes (see phenotype 'Fertilization and ookinete'). Oocysts produce normal numbers of sporozoites that are not affected in their infectivity to salivary glands. Motility of sporozoites was normal. Infectivity of sporozoites as measured by infection of rats (Wistar) by intravenous inoculation of sporozoites was reduced (~1/20 to ~1/50 of wild type sporozoites). Infectivity to HepG2 cells in vitro was not affected (similar numbers of EEFs were formed). However, cell traversal (cell-passage) was strongly impaired (as measured by the 'cell wound assay'). Infectivity of sporozoites to rats (Wistar) was restored in Kupffer cell-depleted rats. |
Liver stage | Infectivity of sporozoites as measured by infection of rats (Wistar) by intravenous inoculation of sporozoites was reduced (~1/20 to ~1/50 of wildtype sporozoites). Infectivity to HepG2 cells in vitro was not affected (similar numbers of EEFs were formed). However, cell traversal (cell-passage) was strongly impaired (as measured by the 'cell wound assay'). Infectivity of sporozoites to rats (Wistar) was restored in Kupffer cell-depleted rats. |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1432300 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1216600 | ||||||||||||||||||||||||
Gene product | cell traversal protein for ookinetes and sporozoites | ||||||||||||||||||||||||
Gene product: Alternative name | CelTOS; cell traversal protein for ookinetes and sporozoites | ||||||||||||||||||||||||
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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 | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Complete | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | pbdhfr | ||||||||||||||||||||||||
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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