| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene disruption
|
| Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 29338985
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| 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 TRAP/FlpL
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| Other information parent line | This line expresses FlpL recombinase under the control of the TRAP promoter, active in the mosquito oocyst stage, and selectively excises the DNA sequence flanked by FRT sites (S. Mishra, K.A. Kumar and P. Sinnis, unpublished data). The FlpL expression cassette is introduced into the silent p230p gene locus (by GIMO transfection using line 1596cl1; RMgm-687) resulting in line 1809 (RMgm-4186). The line has been generated in Leiden and cloned in the Sinnis lab. This line does not contain a drug-selectable marker. |
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| The mutant parasite was generated by |
| Name PI/Researcher | Choudhary HH, Mishra S |
| Name Group/Department | Division of Parasitology |
| Name Institute | CSIR-Central Drug Research Institute |
| City | Lucknow |
| Country | India |
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| Name of the mutant parasite |
| RMgm number | RMgm-4412 |
| Principal name | Cs mutant |
| Alternative name | |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | Yes |
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| Phenotype |
| Asexual blood stage | Not different from wild type |
| Gametocyte/Gamete | Not different from wild type |
| Fertilization and ookinete | Not different from wild type |
| Oocyst | Not different from wild type |
| Sporozoite | Not different from wild type |
| Liver stage | Not different from wild type |
| Additional remarks phenotype | Mutant/mutation
The mutant lacks expression of Chorismate synthase (Cs). The mutant was obtained after excision of the cs gene by the Flp/FRT system. The mutant expresses Flp recombinase under the control of the promoter of trap. The cs gene has been removed from the genome by using the Flp/FRT site-specific recombination (SSR) system. Removal of the cs gene has been achieved by transmission of the mutant through mosquitoes, thereby activating expression of the FlpL recombinase that resulted in the excision of the cs gene which was flanked by FRT sequences. After mosquito passage parasites were cloned and cloned mutants were selected and analysed that lack the cs gene
Protein (function)
The shikimate pathway has seven steps that finally lead to the formation of chorismate from 5-enolpyruvylshikimate 3-phosphate (EPSP). Chorismate produced as the end product of this pathway serves as a substrate for the synthesis of aromatic amino acids, Para-aminobenzoic acid (PABA) and ubiquinones. PABA is a precursor for folates that are required for nucleotide synthesis and the methylation cycle. Ubiquinones are components of the electron transport chain, and in Plasmodium it is synthesised in two steps – the isoprene side chain synthesis occurs via a terpenoid pathway, and benzoquinone synthesis occurs via a shikimate pathway. Chorismate synthase (Cs) is the immediate and only enzyme of this pathway required for chorismate synthesis. CS carries out 1, 4 trans elimination of the phosphate group of EPSP that results in the formation of chorismate, utilising Flavin mononucleotide (FMN) as a cofactor.
Phenotype
No phenotypes were detected in blood, mosquito and liver stages that were different from wild type.
Additional information
Other mutants |
*RMgm-4412
