SummaryRMgm-72
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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) : 11927543 |
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 | V. Thathy, V. Nussenzweig, R. Menard |
Name Group/Department | Michael Heidelberger Division of Immunology |
Name Institute | New york University Medical Center |
City | New York |
Country | USA |
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Name of the mutant parasite | |
RMgm number | RMgm-72 |
Principal name | PINA270-1; PINA270-2 |
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 oocysts are produced in Anopheles stephensi mosquitoes. Sporozoite formation within the oocysts is affected. Oocysts display intermediary levels of extension of the inner membranes and microtubules and at least 50% of the oocyst-sporozoites had abnormal and variable sizes and shapes. |
Sporozoite | The (aberrantly shaped) sporozoites did not invade salivary glands as shown by the greatly reduced ratio of salivary gland to midgut sporozoites and did not show normal gliding motility. Sporozoites were not infective. |
Liver stage | See the description of the phenotype of the sporozoite. |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype The phenotype analyzes of this mutant and the mutant lacking expression of CS (RMgm-9)demonstrate the essential role of CS in the formation of sporozoites within the oocyst. The results of these analyzes indicate that CS is essential for establishing polarity in oocysts by affecting the development of the inner membrane and associated microtubules underneath the oocyst outer membrane, which normally demarcate focal budding sites of daughter cells. 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 | truncation of the 3'UTR of the gene | ||||||||||||||||||||||||||
Inducable system used | No | ||||||||||||||||||||||||||
Short description of the conditional mutagenesis | Not available | ||||||||||||||||||||||||||
Additional remarks inducable system | |||||||||||||||||||||||||||
Type of plasmid/construct | Plasmid single 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 insertion construct used results in introduction replacement of the wild type cs gene with a copy of the cs gene that is under control of the wild type 5'UTR regulatory sequence but contains a truncated 3'UTR sequence. Cloning and sequencing of CS transcripts in P.berghei sporozoites have revealed numerous 3' ends. The 3' ends of the CS transcripts are heterogeneous, located between nucleotides 88 and 337 after the CS stop codon. The majority of the CS 3' ends are clustered 300 bp past the stop codon, just upstream of a 16 bp GU-rich element. In the mutant the CS gene is followed by only 270 bp of its 3'-UTR. The mutant sporozoites reproducibly contained 5x less CS protein than wild-type sporozoites. They still produced a normal ratio of the 44 and 54 kDa forms of CS, which are thought to represent the surface molecule and intracellular precursor of CS, respectively. | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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