Mutant/mutation
In the mutant GFP is expressed under control of the PBANKA_1003900 promoter. The GFP-expression cassette has been introduced as episomes/circular plasmid
This gene is a 'syntenic ortholog' of sexual stage-specific protein precursor (Pfs16; PF3D7_0406200) of P. falciparum.
The phenotype has not been analysed in detail. Very weak (background?) fluorescence was observed in gametocytes.
The similarity of P. falciparum Pfs16 and the syntenic P. berghei gene (Pbs16?) is below threshold.
See also RMgm-1439 for a mutant with a disrupted PBANKA_1003900) gene locus
See the link PBANKA_1003900 for other mutants
Additional information from the paper
PBANKA_1003900 was found to be highly transcribed in liver stages.
PBANKA_1003900 is a syntenic ortholog of P. falciparum sexual stage-specific protein precursor (Pfs16; PF3D7_0406200) which is expressed early during development of P. falciparum gametocytes. Plasmodium berghei parasites expressing an mCherry-tagged PBANKA_1003900 provided experimental evidence that this gene is also expressed in gametocytes and was, therefore, annotated as a gametocyte specific protein. However, substantial PBANKA_1003900 transcript levels were only detected in liver stages.
Mutants lacking expression of PBANKA_1003900 show no phenotype throughout the complete life cycle, including liver stages.
GFP was not detectable in any of the blood stages, including gametocytes. In contrast, GFP expression was detected by fluorescence microscopy of infected HeLa cells fixed at different time points (24 h, 48 h, 54 h, 60 h). At 24 h of liver/EEF stage development no or only very weak GFP-fluorescence was detectable. At 48 h of EEF stage development, the signal was readily visible and at 54 h and 60 h post infection the fluorescent signal was profoundly intense. When performing live cell imaging, the first signals were observed at 30 h post infection. From 45 h onwards the protein was substantially expressed confirming the results obtained from fixed cells. These fluorescence patterns (fixed and live) nicely confirmed the RNA-seq data during EEF stage development. Interestingly, analyses of gene-deletion mutants lacking PBANKA_1003900 demonstrated that the gene is not essential at any developmental stage throughout the complete parasite life cycle. According to the expression profile deduced from the RNA-seq analysis and also confirmed by the promoter analyses, it is appropriate to revise the annotation as “gametocyte specific protein” and to rename it as “liver specific protein 3 (LISP3)”. |