RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-5585
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Last modified: 27 November 2024, 19:00
*RMgm-5585| 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 | 4 |
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 38888125 |
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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 | Not applicable |
| Other information parent line | |
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| Attempts to generate the mutant parasite were performed by | |
| Name PI/Researcher | Chatterjee T, Habib S |
| Name Group/Department | Department of Biochemistry and Molecular Biology |
| Name Institute | CSIR-Central Drug Research Institute |
| City | Lucknow |
| Country | India |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_0301700 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_0203900 | ||||||||||||||||||||||||
| Gene product | 5'-3' exonuclease, putative | ||||||||||||||||||||||||
| Gene product: Alternative name | PbExo | ||||||||||||||||||||||||
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| Details of the genetic modification | |||||||||||||||||||||||||
| Inducable system used | No | ||||||||||||||||||||||||
| Additional remarks inducable system | |||||||||||||||||||||||||
| Type of plasmid/construct used | (Linear) 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 | Complete | ||||||||||||||||||||||||
| 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 | PbExo N-terminal region is predicted to carry an apicoplast targeting sequence. The unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood stage growth/multiplication. The attempts to disrupt PbExo (PBANKA_0301700) were performed with a replacement plasmid containing GFP reporter and hDHFR:yFCU selection marker. Two 5 - and 3'-UTR homology regions F1 and F2 (0.57Kb) were amplified using primers 1430/1431 and 1432/1433 and cloned at SalI and NotI/AscI respectively in the Pb C-GFP-hDHFR:yFCU vector. The construct was linearized with XhoI/AscI and transfected into P. berghei | ||||||||||||||||||||||||
| 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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