Mutant/mutation
The mutant lack expression of UIS4 (up-regulated in infective sporozoites gene 4) and expresses RFP under the control of the constitutive eef1a promoter.
A two step PCR technique was adapted for the efficient generation of the targeting construct. See figure 1: Mikolajczak et.al., Mol Biochem Parasitol 158 (2007) 213-216. In this paper a cloning procedure is described for gene replacement by double homologous recombination in P. yoelii, which requires only one digestion and ligation step. This significantly shortens the time required to complete the production of the targeting vector. Furthermore, for more efficient phenotypic evaluation of the mutant parasites, a fluorescent protein (RFP) cassette is introduced into the targeting vector in which RFP is under the control of the constitutive eef1a promoter. This allows for a more rapid assessment of parasite growth in all of its developmental stages. In addition, the introduction of the fluorescent marker via the replacement strategy confers the stable integration of the marker.

Protein (function)
UIS4 has been identified by transcription-profiling of sporozoites (Kaiser et al., (2001). Mol. Microbiol. 51, 1221-32). The protein is expressed throughout liver stage development and localizes to the parasitophorous vacuole membrane.

Phenotype
See the mutants RMgm-35, RMgm-38, RMgm-39 for a description of the phenotype of mutants lacking expression of UIS4. This paper does not describe the phenotype of the parasites, only the method of generation of the parasites (see 'Mutant/mutation').

Additional information

Other mutants
Other mutants lacking expression of UIS4: RMgm-35, RMgm-38, RMgm-39