Back to search resultsSummaryRMgm-68
|
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) : 16201021 |
MR4 number | |
top of page | |
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 | |
top of page | |
The mutant parasite was generated by | |
Name PI/Researcher | Q. Wang, V. Nussenzweig |
Name Group/Department | Department of Pathology |
Name Institute | Michael Heidelberger Division, New York University School of Medicine |
City | New York |
Country | USA |
top of page | |
Name of the mutant parasite | |
RMgm number | RMgm-68 |
Principal name | CS-RIImut |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
top of page | |
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. After day 14 oocyst-sporozoite numbers remain high compared to wild type, whereas in the hemolymph only minimal numbers of free sporozoites were detected, indicating a defect in release of sporozoites in the hemolymph. Mutant oocysts were more resistant to trypsin treatment (see further additional remarks phenotype). |
Sporozoite | See also the description of the phenotype of the oocyst. Mutant sporozoites display normal morphology and motility. Oocyst-sporozoites obtained by disruption of oocysts are not infective to young Sprague/Dawley rats and they show a reduced binding rate (~40%) to HepG2 cells in vitro. |
Liver stage | Oocyst-sporozoites obtained by disruption of oocysts are not infective to young Sprague/Dawley rats and they show a reduced binding rate (~40%) to HepG2 cells in vitro. |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype Other mutants
|
top of page | |||||||||||||||||||||||||||
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 | ||||||||||||||||||||||||||
top of page | |||||||||||||||||||||||||||
Details of the genetic modification | |||||||||||||||||||||||||||
Short description of the mutation | Region II plus (substituted with alanines) | ||||||||||||||||||||||||||
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 mutant expresses CS in which region II-plus of the protein has been substituted with alanines (R290A, K291A, R292A, K293A). Region II-plus is located at the 5' end of the thrombospondin type I repeat (TSR) domain. Mutations were introduced into the CS coding region by using QuikChange Site-Directed Mutagenesis Kit (Stratagene, La Jolla, California, United States). Primer1 (sense, 5′-GTATAAGAGTTGCTGCAGCAGCAGGTTCAAATAAGAAAGC-3′) and its reverse and complement primer2 were used to mutate R290, K291, R292, and K293 to alanines. | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
| |||||||||||||||||||||||||||
top of page |