RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-544
|
||||||||||
Last modified: 27 May 2012, 17:20
*RMgm-544| 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) : 20951971 |
| top of page | |
| 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). |
| top of page | |
| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | R. Tewari, O. Billker |
| Name Group/Department | University of Nottingham |
| Name Institute | University of Nottingham |
| City | Nottingham |
| 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 | ||||||||||||||||||||||||
| top of page | |||||||||||||||||||||||||
| 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 | nearly complete | ||||||||||||||||||||||||
| 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 | The gene has been targeted for gene deletion using a construct aimed at integration into the genome by double cross-over homologous recombination. The unsuccessful attempts to disrupt this gene indicates an essential function during blood stage development. The gene has been targeted for disruption in a 'kinome-wide' study for deletion of genes encoding Plasmodium protein kinases (protein kinase-like proteins). See the paper for additional information See also: RMgm-325: Unsuccessful attempts to disrupt the pkg gene RMgm-311: A mutants expressing a GFP-tagged (C-terminal) form of PKG 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 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 | |||||||||||||||||||||||||
|
Primer information: Primers used for amplification of the target sequences
 Primer information: Primers used for amplification of the target sequences
| |||||||||||||||||||||||||
| top of page | |||||||||||||||||||||||||