| 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) : 20886115
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| 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 | U. Straschil; R. Tewari |
| Name Group/Department | Institute of Genetics, School of Biology |
| Name Institute | University of Nottingham |
| City | Nottingham |
| Country | UK |
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| Name of the mutant parasite |
| RMgm number | RMgm-589 |
| Principal name | PbPF16-GFP |
| Alternative name | |
| Standardized name | |
| Is the mutant parasite cloned after genetic modification | Yes |
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| Phenotype |
| Asexual blood stage | Not different from wild type |
| Gametocyte/Gamete | Faint, cytoplasmic, GFP expression in the male gametocytes. After induction of male gamete formation, GFP was associated with growing cytoplasmic axonemes (as detected by a-tubulin immunostaining). When male gametes were emerging from the residual body of the male gametocyte, PF16-GFP was localised to the emerging male gamete and had patchy distribution along the length of the gamete. The protein was not detected in female gametocytes and in activated female gametes or in the ookinete. |
| Fertilization and ookinete | Faint, cytoplasmic, GFP expression in the male gametocytes. After induction of male gamete formation, GFP was associated with growing cytoplasmic axonemes (as detected by a-tubulin immunostaining). When male gametes were emerging from the residual body of the male gametocyte, PF16-GFP was localised to the emerging male gamete and had patchy distribution along the length of the gamete. The protein was not detected in female gametocytes and in activated female gametes or in the ookinete. |
| Oocyst | Not tested |
| Sporozoite | Not tested |
| Liver stage | Not tested |
| Additional remarks phenotype | Mutant/mutation
The mutant expresses a GFP-tagged form (C-terminal) of PF16 (PBANKA_091740; PF11_0318)
Note that the PF16 gene is distinct from Pfs16, another gametocyte specific gene identified in P. falciparum (PFD0310w).
Protein (function)
All eukaryotic flagella contain an axoneme, a structure consisting of a central apparatus (a pair of microtubules named C1 and C2) encircled by nine doublet microtubules. PF16 is an important Armadillo (ARM)-repeat protein of the central apparatus of eukaryotic flagella. ARM-repeats consist of a ~42-amino acid structurally conserved repeating motif named after the Drosophila segment polarity gene Armadillo (mammalian homologue, b-catenin). Proteins containing ARM-repeats have diverse roles in eukaryotes, including cell signalling, cytoskeletal organisation and regulation of gene expression. Inactivation of the Chlamydomonas PF16 gene led to loss of the central pair of microtubules and abnormal flagellar function, showing that PF16 is required for flagellar motility and stability of the central apparatus C1 microtubule. Disruption of SPAG6, the mouse PF16 orthologue, results in male infertility, due to loss of sperm motility.RNAi of PF16 in the flagellated protozoan Trypanosoma brucei demonstrates a conserved role in flagellum-dependent motility.
Phenotype
The analyses indicate that PF16 is expressed in male gametocytes and male gametes.
See also RMgm-588 for a mutant lacking expression of PF16. Phenotype analyses of this mutant indicate that PF16 plays a crucial role maintaining the correct microtubule structure in the central apparatus of the axoneme of the male gamete . Mutant male gametes show abnormal flagellar movement and reduced fertility, but lack of PF16 expression does not lead to complete sterility.
Additional information
See also RMgm-588 for a mutant lacking expression of PF16. Phenotype analyses of this mutant indicate that PF16 plays a crucial role maintaining the correct microtubule structure in the central apparatus of the axoneme of the male gamete . Mutant male gametes show abnormal flagellar movement and reduced fertility, but lack of PF16 expression does not lead to complete sterility.
Other mutants
RMgm-588: a mutant lacking expression of PF16
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*RMgm-589
