RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-325
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Last modified: 21 December 2011, 15:53
*RMgm-325| 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) : 19779564 |
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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 | R.W. Moon; D.A. Baker; O. Billker |
| Name Group/Department | Department of Cell and Molecular Biology |
| Name Institute | Imperial College London |
| City | London |
| Country | UK |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1008200 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1436600 | ||||||||||||||||||||||||
| Gene product | cGMP-dependent protein kinase | ||||||||||||||||||||||||
| Gene product: Alternative name | PKG | ||||||||||||||||||||||||
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| Details of the genetic modification | |||||||||||||||||||||||||
| Inducable system used | No | ||||||||||||||||||||||||
| Additional remarks inducable system | |||||||||||||||||||||||||
| Type of plasmid/construct used | Unknown | ||||||||||||||||||||||||
| 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 | Unknown | ||||||||||||||||||||||||
| Promoter of the selectable marker | Unknown | ||||||||||||||||||||||||
| Selection (positive) procedure | Unknown | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | In the paper it is reported that repeated attempts were made to disrupt the gene. No information/details is provided on the number of attempts to disrupt the gene and the construct used to disrupt the gene. (Gene models for PKG PB000726.02.0; PB300651.00.0) See RMgm-311 for a mutant expressing a GFP-tagged (C-terminal) form of PKG (cGMP-dependent protein kinase). See RMgm-357 for a ‘conditional knock-out mutant’ lacking expression of PKG in liver stages. The Flp/FRT site-specific recombination (SSR) system of yeast has been used to silence PKG expression specifically in liver stages Cyclic GMP-dependent protein kinases (PKGs) are the major mediators of the cGMP signal transduction pathway and regulate a variety of physiological effects. In the paper it is shown that a selective inhibitor of apicomplexan PKG, the ATP analog Compound 1, potently blocked ookinete motility. In the paper evidence is presented that in ookinetes GCß is the main source for cGMP and that a cyclic GMP signaling module exists that regulates gliding motility of ookinetes. This evidence is partly based on the analysis of mutants lacking expression PDEδ (RMgm-308) and GCβ (RMgm-307) and a mutant lacking expression of both GCβ and PDEδ (RMgm-309). These analyses indicate that PDEδ is the relevant cGMP degrading enzyme. Signalling via a cGMP-dependent protein kinase (PKG; PF14_0346; PB000726.02.0) may regulate ookinete differentiation and motility. RMgm-544: Unsuccessful attempts to disrupt the pkg gene RMgm-311: A mutant expressing a GFP-tagged (C-terminal) form of PKG Disruption of the P. falciparum ortholog has been attempted (Solyakov et al., 2011, Nat Commun, 2:565). The gene is likely essential for asexual proliferation as integration of the KO vector was not achieved. Accesibility of the locus to recombination was verified by C-terminal tagging. | ||||||||||||||||||||||||
| 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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