Back to search resultsSummaryRMgm-996
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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 | Unknown |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 24594931 |
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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 2.34 |
Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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Attempts to generate the mutant parasite were performed by | |
Name PI/Researcher | Brochet, M; Billker, O. |
Name Group/Department | Wellcome Trust Sanger Institute, Hinxton |
Name Institute | Wellcome Trust Sanger Institute, Hinxton |
City | Hinxton, Cambridge |
Country | UK |
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Details of the target gene | |||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1109400 | ||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0509800 | ||||||||||||||||||||||||
Gene product | phosphatidylinositol 4-kinase | ||||||||||||||||||||||||
Gene product: Alternative name | PI4K; PI(4)K | ||||||||||||||||||||||||
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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 | Yes | ||||||||||||||||||||||||
Name of PlasmoGEM construct/vector | - | ||||||||||||||||||||||||
Modified PlasmoGEM construct/vector used | No | ||||||||||||||||||||||||
Plasmid/construct map | |||||||||||||||||||||||||
Plasmid/construct sequence | |||||||||||||||||||||||||
Restriction sites to linearize plasmid | |||||||||||||||||||||||||
Partial or complete disruption of the gene | Unknown | ||||||||||||||||||||||||
Additional remarks partial/complete disruption | |||||||||||||||||||||||||
Selectable marker used to select the mutant parasite | hdhfr/yfcu | ||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||
Additional remarks genetic modification | No details are provided for the PlasmoGEM vector used. In the paper allelic replacement mutants are described expressing mutated forms of PI4K: mutated resisue S534 to alanine, either on its own or in combination with a nearby phosphorylation site, S538. At the ookinete stage, pi4kS534A and pi4kS534A/S538A clonal mutants showed a significant decrease in gliding speed compared with a control line, pi4kS534 which would be consistent with phosphorylation of S534 in PI4K contributing to the regulation of phosphoinositide metabolism in vivo. Enzymes in the inositol phospholipid biosynthetic pathway. Phosphorylated phosphatidylinositol lipids have important roles in vesicle trafficking and as a source of secondary messengers in signal transduction. Their biosynthesis from phosphatidyl-1D-myo-inositol (PI) is mediated by lipid kinases. The P. berghei genome encodes four putative lipid kinases to convert PI first to phosphatidylinositol 4-phosphate (PI4P) and then to phosphatidylinositol (4,5)- bisphosphate (PI(4,5)P2). Hydrolysis of the latter by a PI-specific phospholipase C (PI-PLC) gives rise to the secondary messenger inositol (1,4,5)-trisphosphate (IP3), which plays an important role in P. berghei gametocytes, where it is responsible for the mobilisation of Ca2+ from internal stores, leading to activation and gametogenesis. To test whether the PKG-dependent phosphorylation of enzymes associated with phosphoinositide metabolism has a direct role in ookinete motility, an experimental genetics approach was used to infer the role of putative PI kinases. Both the putative PI4K (PBANKA_110940) and the putative PIP5K (PBANKA_020310) were unable to be genetically disrupted (unpublished data), suggesting these genes may be essential for asexual growth, although both loci could be modified | ||||||||||||||||||||||||
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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