SummaryRMgm-35
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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) : 15699336 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei NK65 |
Name parent line/clone | Not applicable |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | A.K. Mueller, K. Matuschewski, 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-35 |
Principal name | uis4REP- |
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. Liver stage numbers are reduced by ~50% twenty four hours after invasion of HepG2 cells. At later time points, mature liver schizonts are absent. Only developmentally arrested liver forms persist. Infection of mice (C57BL/6) by bite of infected mosquitoes or subcutaneous inoculation of sporozoites did not result in blood stage infection. Only intraveneous inoculation of very high numbers of sporozoites (50.000) resulted in blood stage infection in ~50% of the mice. These mice showed a prolonged prepatent period. |
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 | PBANKA_0501200 | ||||||||||||||||||||||||
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; 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 single 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 | The mutant described here, uis4REP-, is generated using a replacement vector resulting in disruption of uis4 through double cross-over recombination. In the same paper (Mueller et al., (2004) PNAS, 102: 3022-37) a mutant (uis4-) has been described that has been generated using a disruption vector, resulting in disruption of uis4 by single cross-over recombination. | ||||||||||||||||||||||||
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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