SummaryRMgm-74
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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) : 15839899 |
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 2.34 |
Other information parent line | P. berghei ANKA 2.34 is a cloned, gametocyte producer line of the ANKA strain (PubMed: PMID: 15137943). |
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The mutant parasite was generated by | |
Name PI/Researcher | R. Tewari, D. Rathore, A. Crisanti |
Name Group/Department | Imperial College of Science, Technology and Medicine |
Name Institute | Imperial College of Science |
City | London |
Country | UK |
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Name of the mutant parasite | |
RMgm number | RMgm-74 |
Principal name | PgCS-14 |
Alternative name | PgCS-14 (replacement with PgCS) |
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 | Normal numbers of of mature oocysts and oocyst-sporozoites at day 14 after infection of A. stephensi mosquitoes. Oocysts and oocyst-sporozoite numbers in A. gambiae were comparable to that of wild type. Both wild type and mutant parasites did not infect Aedes aegypti. A proportion (12%) of PgCS-14 infected A. stephensi mosquito, showed melanotic encapsulation of oocysts at late (18–21 days p.i.) developmental stages. |
Sporozoite | Normal numbers of oocyst-sporozoites are formed but sporozoites fail to invade salivary glands. The frequency of motile oocyst-derived sporozoites was comparable to that observed for oocyst-derived sporozoites of wild type. Sporozoites were not infective to mice (C57Bl/6). |
Liver stage | Sporozoites were not infective to mice (C57Bl/6). |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype Other mutants |
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0403200 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0304600 | ||||||||||||||||||||||||||
Gene product | circumsporozoite (CS) protein | ||||||||||||||||||||||||||
Gene product: Alternative name | CSP | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Short description of the mutation | The endogenous P. berghei gene replaced with the homolog of P. gallinaceum | ||||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||||
Short description of the conditional mutagenesis | Not available | ||||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||||
Type of plasmid/construct | 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 | tgdhfr | ||||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | The complete CS coding sequence from P. gallinaceum was introduced into the genome, thereby replacing the endogenous P. berghei CS gene. The pgCS gene is under control of the PbCS regulatory sequences. In the paper details are missing (not provided!) on the sequence of the CS gene of P. gallinaceum and no details are given of the primers used for the target sequences to introduce the construct into the genome | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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