SummaryRMgm-4658
|
Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) | Gene tagging |
Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 31370870 |
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 | |
top of page | |
The mutant parasite was generated by | |
Name PI/Researcher | Yeoh LM, McFadden GI, Ralph SA |
Name Group/Department | Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institut |
Name Institute | The University of Melbourne |
City | Parkville |
Country | Australia |
top of page | |
Name of the mutant parasite | |
RMgm number | RMgm-4658 |
Principal name | PbSR-MG HA-tagged |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
top of page | |
Phenotype | |
Asexual blood stage | Localizing an integrated, hemagglutinin-tagged (HA-tagged) PbSR-MG confirmed its expected nuclear localization |
Gametocyte/Gamete | Localizing an integrated, hemagglutinin-tagged (HA-tagged) PbSR-MG confirmed its expected nuclear localization |
Fertilization and ookinete | Not tested |
Oocyst | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation Localizing an integrated, hemagglutinin-tagged (HA-tagged) PbSR-MG confirmed its expected nuclear localization Phenotype Phenotype analyses of a mutant lacking expression of PbSR-MG showed the follwing (see mutant RMgm-4657):
|
top of page | |||||||||||||||||||||||||||
Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_0921600 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_1126800 | ||||||||||||||||||||||||||
Gene product | RNA-binding protein, putative | ||||||||||||||||||||||||||
Gene product: Alternative name | PbSR-MG | ||||||||||||||||||||||||||
top of page | |||||||||||||||||||||||||||
Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | triple-HA | ||||||||||||||||||||||||||
Details of tagging | C-terminal | ||||||||||||||||||||||||||
Additional remarks: tagging | |||||||||||||||||||||||||||
Commercial source of tag-antibodies | |||||||||||||||||||||||||||
Type of plasmid/construct | (Linear) 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 | hdhfr | ||||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | The hemagglutinin epitope-tagged line was created by modification of a pL0006-based plasmid. This plasmid was designed for single-site recombination into P. berghei and includes three hemagglutinin (HA) tags, followed by the 3'-UTR of KAHRP, followed by a DHFR selectable marker. We cloned the 3' end of PbSR-MG from genomic DNA with the primers “PbSR-MG 3'-HindIIIApaI SP” and “PbSR-MG 3_-AvrII-BglII ASP” and ligated this into the vector backbone. | ||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
| |||||||||||||||||||||||||||
top of page |