RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-34
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Last modified: 18 October 2018, 12:05
*RMgm-34| 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) : 17418435 |
| 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 | M. Labaied, 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-34 |
| Principal name | Pbtrsp(−) |
| 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 | Normal numbers of salivary gland sporozoites are produced. Sporozoites show a gliding motility similar to wild type sporozoites. Invasion of hepatocytes is significantly reduced (see phenotype 'liver stage'). |
| Liver stage | In vitro invasion of hepatocytes by the sporozoites is significantly reduced (35-38% reduction in number of completely intracellular sporozoites compared to wild type sporozoites). Invaded sporozoites develop normally inside HepG2 cells into mature liver schizonts. In vivo a 40-80% reduction in liver infection is observed after infection of C57BL/6 mice by intraveneous inoculation of sporozoites. |
| Additional remarks phenotype | The mutant lacks expression of the thrombospondin related sporozoite protein (TRSP), a thrombospondin type 1 repeat-containing protein (TRS, TSP1). The phenotype analysis indicates a role of this protein in heptocyte entry. |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0209100 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0104000 | ||||||||||||||||||||||||
| Gene product | thrombospondin-related sporozoite protein | sporozoite stage TSP1 domain protein | ||||||||||||||||||||||||
| Gene product: Alternative name | thrombospondin related sporozoite protein; TRSP; Sporozoite stage TSP1 domain protein, S21 | ||||||||||||||||||||||||
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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 | |||||||||||||||||||||||||
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Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
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