RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-389
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Last modified: 18 February 2010, 18:56
*RMgm-389| 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) : 19000816 |
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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 | P. berghei ANKA cl15cy1 |
| Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255) |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | I. Kursula; H. Schüler |
| Name Group/Department | Department of Biochemistry |
| Name Institute | University of Oulu |
| City | Oulu |
| Country | Finland |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0833000 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0932200 | ||||||||||||||||||||||||
| Gene product | profilin | ||||||||||||||||||||||||
| Gene product: Alternative name | Pfn | ||||||||||||||||||||||||
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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 | SacII, KpnI | ||||||||||||||||||||||||
| 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 | No detailed information on primer sequences is provided. Attempts to disrupt the profilin gene of P. berghei were unsuccessful suggesting an essential role during blood stage development. Profilins are ubiquitous and essential actin monomer binding proteins, known to interact with proline-rich regions in a variety of proteins and to regulate actin filament formation. For fast actin polymerization at selected sites, profilin is an essential control element by recruiting actin monomers in a polymerizable form to actin polymerization machineries. Profilins are usually defined by their shared, highly conserved structure and by their ability to bind actin, proline-rich sequences The genome of the different Plasmodium spcies contains a 'profilin' ortholog In this study it is shown that profilin of P. falciparum adopts an overall core structure and displays biochemical properties that together allow for its classification as a conventional profilin. However, it also possesses a unique minidomain, including an arm-like b-hairpin protrusion common to apicomplexan parasites, and an acidic loop specific for Plasmodium species. Profilin is an essential gene for Plasmodium blood stages. In this study a control replacement plasmid was generated that contained a functional full-length copy of the human profilin gene (Pfn1) under the control of the endogenous PbPfn promoter and a constitutive untranslated region to permit proper transcript stability. Transfection of this construct yielded viable parasites that contained the predicted allelic replacement at the first attempt. These findings show that (i) the PbPfn locus is readily accessible to homologous recombination, and (ii) human profilin can complement the essential role of the parasite protein. | ||||||||||||||||||||||||
| 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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