Back to search resultsSummaryRMgm-4409
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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) : 29307698 |
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 | Deligianni, E; Siden-Kiamos, I. |
Name Group/Department | Institute of Molecular Biology and Biotechnology |
Name Institute | FORTH |
City | Heraklion |
Country | Greece |
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Name of the mutant parasite | |
RMgm number | RMgm-4409 |
Principal name | 100390-mCherry |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | No |
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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 | Not tested |
Sporozoite | Not tested |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation To investigate protein expression a construct encoding PBANKA_1003900 fused at the C-terminus to mCherry was integrated by single cross-over in the endogenous locus and a clonal line was generated (the mCherry-tagged gene is under control of the 3'UTR of the PPPK-DHPS gene (used in the pBAT-SIL6 vector). Preliminary experiments revealed mCherry signal in the periphery of gametocytes Immunofluorescence analysis using an anti-mCherry antibody detected a diffuse signal in the cytoplasm of the parasite with no clear localization to the PVM (data not shown). We were thus unable to definitely localize the protein to the PVM. Western blot analysis revealed that a protein of the expected molecular weight was present in gametocytes. In ookinetes samples a very weak band was only seen after exposure for a very long time compared to the strong signal of the gametocyte sample at short exposure time. Most likely these traces of protein are derived from gametocytes persisting in the ookinete culture. See also RMgm-968 for a mutant with a disrupted PBANKA_100390) gene locus
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Details of the target gene | |||||||||||||||||||||||||||
Gene Model of Rodent Parasite | PBANKA_1003900 | ||||||||||||||||||||||||||
Gene Model P. falciparum ortholog | PF3D7_0406200 | ||||||||||||||||||||||||||
Gene product | sexual stage-specific protein precursor | ||||||||||||||||||||||||||
Gene product: Alternative name | Pfs16 | ||||||||||||||||||||||||||
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Details of the genetic modification | |||||||||||||||||||||||||||
Name of the tag | mCherry | ||||||||||||||||||||||||||
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/yfcu | ||||||||||||||||||||||||||
Promoter of the selectable marker | eef1a | ||||||||||||||||||||||||||
Selection (positive) procedure | pyrimethamine | ||||||||||||||||||||||||||
Selection (negative) procedure | No | ||||||||||||||||||||||||||
Additional remarks genetic modification | |||||||||||||||||||||||||||
Additional remarks selection procedure | |||||||||||||||||||||||||||
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