Back to search resultsSummaryRMgm-73
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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) : 16207248 |
MR4 number | |
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Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei NK65 |
Name parent line/clone | Not applicable |
Other information parent line | |
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The mutant parasite was generated by | |
Name PI/Researcher | Q. Wang, H. Fujioka, V. Nussenzweig |
Name Group/Department | Department of Pathology |
Name Institute | Michael Heidelberger Division, New York University School of Medicine |
City | New York |
Country | USA |
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Name of the mutant parasite | |
RMgm number | RMgm-73 |
Principal name | CS-∆GPI, CS-TM1, CS-TM2 |
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 | Normal numbers of of mature oocysts are produced by all three mutants in A. stephensi mosquitoes. However no sporozoites are formed within the oocysts. Up to day eight after feeding of the mosquitoes the morphology of oocysts is normal; after day 8, the oocysts display a highly vacuolated structure (see for further details 'Additional remarks phenotype'). |
Sporozoite | See phenotype description of the oocyst and for further details 'Additional remarks phenotype'. |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype The phenotype analysis shows that normal numbers of mature oocysts are produced by all three mutants in A. stephensi mosquitoes. However no sporozoites are formed, comparable to the phenotype of a mutant lacking CS (RMgm-9) with a comparable (EM) morphology of the aberrant oocysts. These results suggest that CS function during sporozoite development occurs on the plasma membrane of the oocysts and requires the specific presence of its GPI-anchor. 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 | Mutations of the C-terminal glycosylphosphatidylinositol (GPI)-anchor addition sequence | ||||||||||||||||||||||||||
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 | pbdhfr | ||||||||||||||||||||||||||
Promoter of the selectable marker | pbdhfr | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | In the mutants the endogenous CS is replaced with the mutated forms using replacement vectors. In CS-∆GPI, the addition of a GPI-anchor to CS is prevented by deletion of nucleotides encoding the C-terminal stretch of hydrophobic amino acids that contains the GPI-anchor addition sequence (S317 to F340). In CS-TM1 and CS-TM2, the GPI-anchor addition sequence of CS was replaced with two versions of the C-terminal of another sporozoite surface protein, TRAP (PB000374.03.0). CS-TM1 contains only the TRAP Trans Membrane Domain (TMD; I544-G559) followed by five alanines at the C-terminus. CS-TM2 contains a larger portion of TRAP (K534-S569), which includes its TMD, 10 amino acids upstream of the TMD (K534–K543) and 10 amino acids downstream from the TMD (V560-S569). The choice of TM2 replacement was based on the finding that these amino acids are sufficient to direct TRAP to the sporozoite plasma membrane | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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