RMgmDB - Rodent Malaria genetically modified Parasites
SummaryRMgm-5125
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Last modified: 20 January 2023, 14:17
*RMgm-5125| 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) : 36657610 |
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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 | Tanneru N, Sijwali PS |
| Name Group/Department | CSIR-Centre for Cellular and Molecular Biology |
| Name Institute | CSIR-Centre for Cellular and Molecular Biology |
| City | Hyderabad |
| Country | India |
Disrupted: Mutant parasite with a disrupted gene| top of page | |||||||||||||||||||||||||
| Details of the target gene | |||||||||||||||||||||||||
| Gene Model of Rodent Parasite | PBANKA_1033200 | ||||||||||||||||||||||||
| Gene Model P. falciparum ortholog | PF3D7_1409300 | ||||||||||||||||||||||||
| Gene product | DNA damage-inducible protein 1, putative | ||||||||||||||||||||||||
| Gene product: Alternative name | |||||||||||||||||||||||||
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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 | ||||||||||||||||||||||||
| Promoter of the selectable marker | unknown | ||||||||||||||||||||||||
| Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||
| Selection (negative) procedure | No | ||||||||||||||||||||||||
| Additional remarks genetic modification | Published in: bioRxiv preprint doi: https://doi.org/10.1101/2021.10.29.466443 The unsuccessful attempts to disrupt this gene indicate an essential function during asexual blood stage growth/multiplication. The PbDDI1 gene was targeted for knock-out and knock-down using double cross-over homologous recombination approach. For the knock-out constructs, the 5'-UTR (flank 1) and 3’-UTR (flank 2) of PbDDI1 were amplified from P. berghei genomic DNA using PbDdi1- Fl1F/ PbDdi1-Fl1R and PbDdi1-Fl2F/PbDdi1-Fl2R primer sets, respectively. The flank 1 and flank 2 were cloned into the HB-DJ1KO plasmid at NotI-KpnI and AvrII-KasI sites, respectively, to obtain HB-PbDDI-(FL1+FL2) plasmid. The GFP coding sequence was excised from pGT-GFPbsc with KpnI-XhoI and subcloned into the similarly digested HB-PbDDI- (FL1+FL2) to obtain HB-pbDDIKO plasmid. | ||||||||||||||||||||||||
| 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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