RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-264
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Last modified: 29 June 2010, 14:08
*RMgm-264| 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) : 20580650 |
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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. Siden-Kiamos, C. Louis |
| Name Group/Department | Institute of Molecular Biology and Biotechnology |
| Name Institute | Foundation for Research and Technology – Hellas, Vassilika Vouton |
| City | Heraklion, Crete |
| Country | Greece |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0209300 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0103800 | ||||||||||||||||||||||||
| Gene product | actin-related protein | ||||||||||||||||||||||||
| Gene product: Alternative name | ARP1; actin3 | ||||||||||||||||||||||||
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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 | |||||||||||||||||||||||||
| Partial or complete disruption of the gene | Partial | ||||||||||||||||||||||||
| 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 | Disruption experiments were performed in the Leiden Malaria Research Group (Exp. 250, 307, 315; plasmid JR16, obtained from I. Siden-Kiamos) and by K. Matuschweski. Attempts to delete the gene in the parasite were unsuccessful, suggesting that ARP1 is essential during blood stage development. Transcription of arp1 was shown to be upregulated in ookinetes compared to gametocytes and asexual blood stages. Western analysis using antiserum against the recombinant protein expressed in bacteria, showed expression in all stages. Actin-related proteins (Arp’s) have a high degree of sequence similarity to conventional actins, their three dimensional structure also being conserved. However, their functions are varied and distinct from that of conventional actins. To date 11 distinct subfamilies of Arp’s from a wide range of eukaryotic organisms have been described, each subfamily having a distinct function. Arp2 and Arp3 are absent in apicomplexan parasites. Arp’s 4-9 have a function in the nucleus; in chromatin remodelling, histone modification and regulation of DNA repair. Only one Arp, namely Arp1, has been shown to form a filamentous homopolymer, with a uniform size of ~40 nm. In addition, similar to actin, it is regulated by ATP turnover. The Arp1 minifilament is a component of the dynactin complex, which is required for dynein transport on microtubules. Dynactin, in turn, which is a large multi-subunit complex, has a function both in mitosis and in vesicular transport. Arp 11 acts as a capping protein on the pointed end of the Arp1 filament in the dynactin complex, at least in some organisms. Genomic sequence analysis of apicomplexan parasites has suggested that several of the subunits of the dynactin complex are present in these parasites. | ||||||||||||||||||||||||
| 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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