RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-298
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Last modified: 27 June 2009, 16:37
*RMgm-298| Successful modification | The gene/parasite could not be changed/generated by the genetic modification. |
| The following genetic modifications were attempted | Gene disruption |
| Number of attempts to introduce the genetic modification | 3 |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 19165333 |
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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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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | C. Agop-Nersesian; M. Meissner; G. Langsley |
| Name Group/Department | Hygieneinstitut, Department of Parasitology |
| Name Institute | University Hospital Heidelberg |
| City | Heidelberg |
| Country | Germany |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1418900 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1320600 | ||||||||||||||||||||||||
| Gene product | ras-related protein Rab-11A | ||||||||||||||||||||||||
| Gene product: Alternative name | |||||||||||||||||||||||||
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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 | HindIII/EcoRI | ||||||||||||||||||||||||
| Partial or complete disruption of the gene | Partial | ||||||||||||||||||||||||
| Additional remarks partial/complete disruption | Leaving the first small exon | ||||||||||||||||||||||||
| Selectable marker used to select the mutant parasite | hdhfr | ||||||||||||||||||||||||
| Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||
| Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | Rab genes encode a subgroup of small GTP-binding proteins within the ras super-family that regulate targeting and fusion of transport vesicles within the secretory and endocytic pathways. In the genome of P. falciparum 11 genes have been identified that encode Rab GTPases. Rab11A is highly conserved in apicomplexan parasites. Parasites expressing GFP-Rab11A throughout the asexual blood cycle (RMgm-299) has been generated to analyse expression in blood stages. This analysis confirmed data from IFA, RT-PCR and microarray data that expression occurs during development of the trophozoite into the schizont and free merozoites. A rather diffuse, vesicular localisation of GFP-Rab11A was found in trophozoites and gametocytes, whereas in schizonts, a clear apical-like localisation was observed. The results indicate that Rab11A has a dynamic localisation during intra-erythrocytic development associating with rhoptries only after their biogenesis. The changing distribution of Rab11A indicate that the GTPase could be performing non-rhoptry associated functions during development of the parasite within red blood cells. Based on studies in both Toxoplasma gondii and P. falciparum evidence is presented in this paper that Rab11A mediated transport might be involved in Inner Membrane Complex (IMC) maturation and IMC interaction with the plasma membrane and also might deliver components of the proto-glidosome to the IMC. It is speculated that Rab11A transports vesicles derived from endosome-like compartments, similar to its known function in other eukaryotes. | ||||||||||||||||||||||||
| 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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