Mutant/mutation
The mutant lacks expression of the RNA binding protein Puf1 and expresses GFP under the constitutive eef1α promoter. This drug-selectable marker hdfr/yfcu has been removed from this mutant by negative selection.

Protein (function)
The roles of Puf (Pumilio and fem-3 mRNA binding factor) proteins are diverse yet intimately involved in the translational regulation of developmental and differentiation factors in organisms as diverse as yeast, C. elegans, Drosophila and humans. Two such proteins, Puf1 (PFE0935c) and Puf2 (PFD0825c), are known in the human malaria parasite, P. falciparum, with orthologs in all Plasmodium spp. characterized. The Plasmodium Puf proteins have the typical highly conserved organization that includes the eight tandem copies of the PUM RNA binding domain (or Pumilio Homology Domain, PHD) at the carboxyterminus of the protein and expectedly PfPuf2 was shown to bind RNA in vitro. In P. falciparum evidence has been reported for a role for Puf2 in gametocyte development although Pfpuf2 is most highly transcribed in sporozoites (see also 'Additional Information').

Phenotype
The phenotype analyses indicate that Puf1 is not essential for blood stage development, production of gametocytes, ookinetes, oocysts and infective sporozoites and for mosquito transmission and liver stage development (see also 'Additional information').

Additional information
Normal numbers of gametocytes and ookinetes are produced. Mutant ookinetes showed a slightly different morphology at the light-microscope level (more 'stumpy and rounder' forms compared to the banana-shaped wild type ookinetes). The morphology of mutant ookinetes have not been analysed in more detail.

See mutant RMgm-515, RMgm-516 and RMgm-591 for P. berghei mutants lacking expression Puf2. These mutants show normal blood stage development, gametocyte, ookinete and oocyst production but sporozoite infectivity is affected.

Mutant RMgm-591 that lacks expression of both Puf1 and Puf2 shows the same phenotype as mutants lacking only expression of Puf2.

In P. falciparum Puf2 expression has been demonstrated in gametocytes and disruption of Pfpuf2 (PFD0825c) resulted in increased gametocyte production and increased differentiation of male gametocytes (Miao J. et al., 2010, 123-1039-1049). The phenotype of the P. falciparum mutants lacking expression of Puf2 has not been analysed during mosquito development.

Other mutants
RMgm-514: An independent mutant lacking expression of Puf1 which expresses GFP under the constitutive eef1α promoter
RMgm-515: A mutant lacking expression of Puf2 (PBANKA_071920)
RMgm-516: An independent mutant lacking expression of Puf2 (PBANKA_071920) which expresses GFP under the constitutive eef1α promoter
RMgm-591: A mutant lacking expression of both Puf1 and Puf2 which expresses GFP under the constitutive eef1α promoter