SummaryRMgm-38
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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) : 17624848 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. yoelii |
Parent strain/line | P. y. yoelii 17XNL |
Name parent line/clone | Not applicable |
Other information parent line | 17XNL is a non-lethal strain of P. yoelii |
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The mutant parasite was generated by | |
Name PI/Researcher | A.S. Tarun, S.H.I. Kappe |
Name Group/Department | Department of Pathobiology |
Name Institute | University of Washington |
City | Seattle |
Country | USA |
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Name of the mutant parasite | |
RMgm number | RMgm-38 |
Principal name | Pyuis4[-] |
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 | Not different from wild type |
Oocyst | Not different from wild type |
Sporozoite | Not different from wild type |
Liver stage | In vitro invasion of hepatocytes by the sporozoites is not affected. Liver stage development is strongly impaired. In vivo (BALB/c mice), the number of liver stages at 2 hours after intravenous injection of sporozoites is reduced by 60%. The number of liver stages at 24h were reduced by >95%. liver stages could not be detected at 40h after infection. Liver stages were morphologically similar to wt liver stages at 6 and 12h after infection, a time when the intrahepatocytic transformation from elongate sporozoite to spherical trophozoites commences. Antibody staining for HEP17, a parasitophorous vacuole membrane (PVM)protein, showed a typical circumferential pattern, which indicates that the PVM is present. Liver stages did not develop into schizonts but appeared to be arrested at the trophozoite stage. Arrested liver stages did not persist in the liver after 40h. Infection of BALB/c mice by intravenous inoculation of high numbers of sporozoites (50.000) did not result in blood stage infection. |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype Additional information Other mutants |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PY17X_0502200 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1016900 | ||||||||||||||||||||||||
Gene product | early transcribed membrane protein 10.3 | protein of early gametocyte 4 | ||||||||||||||||||||||||
Gene product: Alternative name | UIS4, up-regulated in infective sporozoites, ETRAMP10.3 | ||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||
Type of plasmid/construct used | (Linear) 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 | 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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