Back to search resultsSummaryRMgm-71
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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) : 12244064 |
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, 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-71 |
Principal name | CSP(RII-)-7 |
Alternative name | CSP(RII-)-7 (replacement with PfCS without region II) |
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 at day 14 after infection of A. stephensi mosquitoes. |
Sporozoite | Compared to wild type, the mutant showed a 290-fold reduction in the number of salivary gland sporozoites. Sporozoites do not show gliding motility. |
Liver stage | Sporozoites failed to infective mice (C57Bl/6) after intravenous injection of sporozoites or after infection by mosquito bite. |
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 ortholog of P. falciparum, lacking region II | ||||||||||||||||||||||||||
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 | The CS coding sequence(1600bp) from P. falciparum (Wellcome strain)lacking region II was introduced into the P. berghei genome. Deletion of coding region II (WSPCSVTCGNGIQVRIK) was introduced by site-directed mutagenesis. The mutated PfCS coding sequence linked to 250 nucleotides of its 3'-UTR was introduced into the genome, thereby replacing the endogenous P. berghei CS gene. The pfCS gene is under control of the PbCS regulatory sequences: i) a 5'-UTR of the PbCS gene encompassing nucleotides 1-1130 immediately upstream of the start codon and ii) the 3'-UTR encompassing nucleotides 1-1150 downstream of the stop codon. | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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