SummaryRMgm-656
|
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) : 21899701 |
MR4 number | |
top of page | |
Parent parasite used to introduce the genetic modification | |
Rodent Malaria Parasite | P. berghei |
Parent strain/line | P. berghei ANKA |
Name parent line/clone | Not applicable |
Other information parent line | no details of the parasite line are provided |
top of page | |
The mutant parasite was generated by | |
Name PI/Researcher | I. Siden-Kiamos; K. Matuschewski |
Name Group/Department | Institute of Molecular Biology and Biotechnology |
Name Institute | Foundation for Research and Technology- Hellas |
City | Heraklion, Crete |
Country | Greece |
top of page | |
Name of the mutant parasite | |
RMgm number | RMgm-656 |
Principal name | MyoA absent in ookinete (Mao) |
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 | Normal numbers of mature ookinetes are formed. Expression of MyoA was absent in mutant ookinetes. Ookinetes are devoid of motility. Oocyst production was strongly reduced |
Oocyst | Ookinetes are devoid of motility. Oocyst production was strongly reduced |
Sporozoite | See 'Additional remarks phenotype' |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Protein (function) Phenotype Additional information Other mutants |
top of page | |||||||||||||||||||||||||||
Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1355700 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1342600 | ||||||||||||||||||||||||||
Gene product | myosin A | ||||||||||||||||||||||||||
Gene product: Alternative name | MyoA | ||||||||||||||||||||||||||
top of page | |||||||||||||||||||||||||||
Details of the genetic modification | |||||||||||||||||||||||||||
Short description of the mutation | 'promoter-swap' mutant; the promoter of myosin A replaced by the promoter of ama1 (PBANKA_091500) | ||||||||||||||||||||||||||
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 | HpaI | ||||||||||||||||||||||||||
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 | For the 'promoter swap' construct a 1.1 kb 3’ deleted PbMyoA fragment was amplified with primers MyoAfor-BamHI and MyoArev-NdeI. In addition, a 1.3 kb region covering the PbAMA1 promoter was amplified using the primers AMA1for and AMA1rev. A single cross-over event at the site of linearization within the 5' region of the MyoA open reading frame leads to an upstream copy that contains the endogenous promoter before a 3' deleted, non-functional MyoA copy. Downstream of this copy is the positive selectable marker and a functional copy consisting of a full-length MyoA allele, which is under the control of the ama1 promoter. | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
| |||||||||||||||||||||||||||
top of page |