SummaryRMgm-5279
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Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene mutation |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 36810637 |
MR4 number | |
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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 507cl1 (RMgm-7) |
Other information parent line | P.berghei ANKA 507cl1 (RMgm-7) is a reference ANKA mutant line that expresses GFP under the control of a constitutive promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190). |
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The mutant parasite was generated by | |
Name PI/Researcher | Marreiros M, Mota MM |
Name Group/Department | Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina |
Name Institute | Universidade de Lisboa |
City | Lisbon |
Country | Portugal |
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Name of the mutant parasite | |
RMgm number | RMgm-5279 |
Principal name | PbSAMS-DD |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype | |
Asexual blood stage | See below: Additional information |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Protein stabilization in vivo was achieved by providing trimethoprim (TMP) in drinking water of mice for 2 days prior to infection (PbSAMS-DD +TMP), while SAMS conditional knockdown (PbSAMS-DD -TMP) was achieved in non-TMP treated mice, as evidenced by a significant decrease in SAMS-HA-DD protein levels, measured by immunoblotting analysis. Phenotype Other mutants |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0823100 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0922200 | ||||||||||||||||||||||||||
Gene product | S-adenosylmethionine synthetase | ||||||||||||||||||||||||||
Gene product: Alternative name | SAMS | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Short description of the mutation | The sams gene fused to the destabilizing domain (DD). | ||||||||||||||||||||||||||
Inducable system used | DD | ||||||||||||||||||||||||||
Short description of the conditional mutagenesis | See below | ||||||||||||||||||||||||||
Additional remarks inducable system |
![]() ![]() Protein stabilization in vivo was achieved by providing trimethoprim (TMP) in drinking water of mice for 2 days prior to infection (PbSAMS-DD +TMP), while SAMS conditional knockdown (PbSAMS-DD -TMP) was achieved in non-TMP treated mice, as evidenced by a significant decrease in SAMS-HA-DD protein levels, measured by immunoblotting analysis. | ||||||||||||||||||||||||||
Type of plasmid/construct | (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 | |||||||||||||||||||||||||||
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 | The pPbsams.DD.HA construct contains a truncated Pbsams ORF in fusion with a destabilizing domain (DD) – a mutant form of the E. coli dihydrofolate reductase (ecDHFR) protein, engineered to be degraded - an HA tag and also a cassette for transgenic expression of the human dhfr - conferring resistance to pyrimethamine - which is flanked by the sams 3’UTR. | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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