Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) |
Gene tagging
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Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 30625318 |
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 cl15cy1
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Other information parent line | A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255). |
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The mutant parasite was generated by |
Name PI/Researcher | Matz JM, Matuschewski K |
Name Group/Department | Department of Molecular Parasitology, Institute of Biology |
Name Institute | Humboldt University |
City | Berlin |
Country | Germany |
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Name of the mutant parasite |
RMgm number | RMgm-4596 |
Principal name | ACDS-tag |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype |
Asexual blood stage | Transgenic parasite lines expressing the endogenous acds and adcl genes (RMgm-4596, RMgm-4597) fused to mCherry-3xMyc were generated and analyzed by live fluorescence microscopy. Co-localization with a cytosolic GFP marker revealed that both enzymes localize to the parasite cytoplasm. ADCS expression was restricted to blood stage trophozoites and liver stages, but remained below the detection limit in rings, in schizonts, and throughout mosquito infection. This finding indicates distinct roles for ADCS in the mammalian host only. In contrast, ADCL was ubiquitously expressed during life cycle progression. |
Gametocyte/Gamete | Not tested |
Fertilization and ookinete | Not different from wild type |
Oocyst | Not different from wild type |
Sporozoite | Not different from wild type |
Liver stage | Transgenic parasite lines expressing the endogenous acds and adcl genes (RMgm-4596, RMgm-4597) fused to mCherry-3xMyc were generated and analyzed by live fluorescence microscopy. Co-localization with a cytosolic GFP marker revealed that both enzymes localize to the parasite cytoplasm. ADCS expression was restricted to blood stage trophozoites and liver stages, but remained below the detection limit in rings, in schizonts, and throughout mosquito infection. This finding indicates distinct roles for ADCS in the mammalian host only. In contrast, ADCL was ubiquitously expressed during life cycle progression. |
Additional remarks phenotype | Mutant/mutation
The mutant expresses a C-terminal mCherry-3xMyc-tagged version of ACDS
Protein (function)
Malaria parasites replicate at high rates in the mammalian bloodstream. Consequently, they require large quantities of folate and folate precursors, which they scavenge from the serum or synthesize de novo. In contrast, the mammalian host strictly depends on the dietary uptake of preformed folates. Folates promote essential functions during one-carbon transfer reactions and consist of three major moieties: para-aminobenzoate (pABA), pterin, and glutamate.It is believed that chorismate, which is produced via the shikimate pathway, is converted to pABA in a two-step reaction. A potential aminodeoxychorismate synthase (ADCS, PBANKA_0823300) and an aminodeoxychorismate lyase (ADCL, PBANKA_1322100) are predicted by Plasmodium genome sequences.
From the paper: 'We show that the parasite depends on de novo folate synthesis only when dietary intake of pABA by the mammalian host is restricted and that only pABA, rather than fully formed folate, is taken up efficiently. This adaptation, which readily adjusts infection to highly variable pABA levels in the mammalian diet, is specific to blood stages and may have evolved to avoid folate competition between the parasite and its host.'
Phenotype
Transgenic parasite lines expressing the endogenous acds and adcl genes (RMgm-4596, RMgm-4597) fused to mCherry-3xMyc were generated and analyzed by live fluorescence microscopy. Co-localization with a cytosolic GFP marker revealed that both enzymes localize to the parasite cytoplasm. ADCS expression was restricted to blood stage trophozoites and liver stages, but remained below the detection limit in rings, in schizonts, and throughout mosquito infection. This finding indicates distinct roles for ADCS in the mammalian host only. In contrast, ADCL was ubiquitously expressed during life cycle progression.
Other mutants |