Successful modification | The parasite was generated by the genetic modification |
The mutant contains the following genetic modification(s) |
Gene tagging
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Reference (PubMed-PMID number) |
Reference 1 (PMID number) : 19428669 |
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
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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 | M. Tufet-Bayona, C.J. Janse, R.E. Sinden, B. Franke-Fayard |
Name Group/Department | Leiden Malaria Research Group |
Name Institute | Leiden University Medical Center (LUMC) |
City | Leiden |
Country | The Netherlands |
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Name of the mutant parasite |
RMgm number | RMgm-213 |
Principal name | RON2-Myc |
Alternative name | |
Standardized name | |
Is the mutant parasite cloned after genetic modification | Yes |
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Phenotype |
Asexual blood stage | The blood-stage ‘ring-forms’, trophozoites and young schizonts, show no detectable expression of RON2-Myc using monoclonal anti-MYC antibodies. RON2-Myc expression is observed in schizonts and merozoites. Merozoites show a 'two-dot' staining pattern which is characteristic for location in the two rhoptry-organelles. |
Gametocyte/Gamete | Not different from wild type |
Fertilization and ookinete | Not different from wild type |
Oocyst | RON2-Myc is not detected in gametocytes and ookinetes. It is expressed in both oocyst-derived sporozoites, obtained at days 9–13 after infection, and also in salivary gland sporozoites. Staining is observed at the apical end of the sporozoite, extending from the mid portion anterior to the nucleus to the apical tip of the sporozoite. |
Sporozoite | RON2-Myc is expressed in both oocyst-derived sporozoites, obtained at days 9–13 after infection, and also in salivary gland sporozoites. Staining is observed at the apical end of the sporozoite, extending from the mid portion anterior to the nucleus to the apical tip of the sporozoite. |
Liver stage | Not tested |
Additional remarks phenotype | Mutant/mutation
The mutant expresses a c-terminal cMyc-tagged RON2 (rhoptry neck protein 2).
Protein (function)
RON2 shows sequence homology with a proven rhoptry protein of Toxoplasma gondii (145.m00331; twinscan-0698; RON2). See also 'Additional Information'.
Phenotype
The phenotype analyses indicate expression of cMyc-tagged RON2 in rhoptries of merozoites and rhoptries of the sporozoites (see also Additional information'). No expression was observed in ookinetes which is in agreement with the proposed lack of rhoptries in this third invasive form of Plasmodium.
Additional information
In T. gondii RON2 is localized specifically to the duct-like neck portion of the rhoptries, leading to the designation as rhoptry neck (RON) protein. Upon invasion of the host cell, T. gondii RON2 is associated with both the micronemal apical-membrane antigen 1 (AMA1) and RON4, and they localize to the moving junction that is formed upon invasion. The Plasmodium homologue of RON4 has been shown to interact with AMA1 of P. falciparum.
Staining of sporozoites with cMyc-antibodies showed staining at the apical end of the sporozoite, extending from the mid portion anterior to the nucleus to the apical tip of the sporozoite. The staining pattern is comparable to the fluorescence pattern of the GFP-tagged rhoptry protein RAP2/3 in sporozoites (see RMgm-209) which suggests a rhoptry localisation for RON2.
Other mutants
RMgm-214: Unsuccessful attempts to disrupt the ron2 gene |