RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-619
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Last modified: 24 July 2011, 12:10
*RMgm-619| 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) : 21689687 |
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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 | T. Hills; A. Srivastava; AP Waters; J.C. Pizarro |
| Name Group/Department | The SGC (Structural Genomics Consortium) |
| Name Institute | University of Toronto |
| City | Toronto |
| Country | Canada |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0715500 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0413500 | ||||||||||||||||||||||||
| Gene product | phosphoglucomutase-2 | ||||||||||||||||||||||||
| Gene product: Alternative name | PGM2 | ||||||||||||||||||||||||
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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 | HindIII, EcoRI | ||||||||||||||||||||||||
| 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 | PbPGM2 appears to be essential to P. berghei asexual blood stage development by the repeated failure to disrupt the gene. On all three occasions, the mutants failed to grow under selection pressure and integration of the selection cassette at the desired locus was not observed suggesting that PGM2 is essential for asexual stages of the parasite. However, it is possible to tag the protein with GFP at its C-terminus via a single cross over mechanism (see RMgm-620). Two P. falciparum genes code for putative phosphoglycerate mutases (PGMases; PfPGM1 - PF11_0208 and PfPGM2 - PFD0660w), a widespread protein group characterized by the involvement of histidine residues in their catalytic mechanism. Phosphoglycerate mutases (EC 5.4.2.1; PGMase) catalyze the reversible conversion of 2-phosphoglycerate (2-PG) to 3-phosphoglycerate (3-PG).This is an essential component of the glycolysis pathway providing 2-PG to the enzyme enolase and also of the gluconeogenesis pathway where it supplies 3-PG to the phosphoglycerate kinase. Members of the PGM family function as phosphotransferases or phosphohydrolases acting upon a variety of substrates. Aside from metabolic functions, a proportion of PGM family members are involved in cell signaling and/or regulation. In this study the P. berghei homologue of PGM2 (PBANKA_071550)was studied. The P. berghei homolog of PGM1 is PBANKA_092810 | ||||||||||||||||||||||||
| 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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