| Successful modification | The parasite was generated by the genetic modification |
| The mutant contains the following genetic modification(s) |
Gene tagging
|
| Reference (PubMed-PMID number) |
Not published (yet)
|
| 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 |
RMgm-4885
|
| Other information parent line | In this mutant the akarin gene has been deleted. This mutant does not contain a drug selectable marker (SM). The SM has been removed by negative selection |
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| The mutant parasite was generated by |
| Name PI/Researcher | Kehrer J, Frischknecht F |
| Name Group/Department | Center for Infectious Diseases, Integrative Parasitology |
| Name Institute | Heidelberg University Medical School |
| City | Heidelberg |
| Country | Germany |
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| Name of the mutant parasite |
| RMgm number | RMgm-4888 |
| Principal name | akratin-GFP |
| Alternative name | |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | Yes |
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| Phenotype |
| Asexual blood stage | akratin-GFP expression (see below) |
| Gametocyte/Gamete | akratin-GFP expression (see below) |
| Fertilization and ookinete | akratin-GFP expression (see below) |
| Oocyst | akratin-GFP expression (see below) |
| Sporozoite | akratin-GFP expression (see below) |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
In the mutant the endogenous P. berghei akratin gene has been replaced with a P. berghei akratin gene that is C-terminally GFP-tagged. The GFP-tagged akratin gene has been introduced into the akratin locus of a mutant (see RMgm-4885) in which the akratin gene has been deleted.
Published in bioRxiv preprint doi: https://doi.org/10.1101/2020.09.29.318857.
Protein (function)
PbANKA_1105300 was found to be conserved among Plasmodium spp. It shares 82% identity with its orthologue in P. yoelii, but only 35% and 32% with its orthologues in P. falciparum and P. vivax, respectively. In contrast to P. berghei and P. falciparum, the orthologues in P. yoelii and P. vivax are predicted to contain only two TMDs. Furthermore, the P. falciparum protein contains about two times more amino acids than the P. berghei protein. No orthologue was found outside Plasmodium spp
Phenotype
The following evidence is presented on akratin expression and localisation:
'We next investigated the localization of the fluorescent signal in the blood stage and the extracellular forms of the life cycle. This revealed a mostly diffuse signal with some punctae in blood stages of both parasite lines expressing the P. falciparum (see mutant RMgm-4887) or P. berghei akratin-GFP. In non-activated gametocytes the signal was distributed within the gametocyte cytoplasm but it was much weaker in the P. falciparum akratin-GFP line, where it appeared punctate. No difference between male and female gametocytes could be observed. However, upon activation two distinct populations, most likely males and females could be observed. While in females the protein was distributed in the cytoplasm of the cell, male (determined by a cloudy Hoechst staining) showed one bright spot in addition to a few weak dots at the periphery. Ookinetes showed a vesicular/endomembranous staining with a prominent peripheral staining for the P. berghei akratin-GFP but an exclusively vesicular pattern for the P. falciparum akratin- GFP. Sporozoites isolated from the oocysts or salivary glands showed a similar punctate localization for both lines reminiscent of secretory vesicles but distinct from the strong apical signal seen in GFP-TRAP sporozoites
Additional information
Other mutants |
*RMgm-4888
