Translational repression (TR) plays an important role in post-transcriptional regulation of gene expression and embryonic development in metazoans. TR also regulates the expression of a subset of the cytoplasmic mRNA population during development of fertilized female gametes of the unicellular malaria parasite, Plasmodium spp. which results in the formation of a polar and motile form, the ookinete. We report the conserved and sex-specific regulatory role of either the 3'- or 5'-UTR of a subset of translationally repressed mRNA species as shown by almost complete inhibition of expression of a GFP reporter protein in the female gametocyte. A U-rich, TR-associated element, identified previously in the 3'-UTR of TR-associated transcripts, played an essential role in mediating TR and a similar region could be found in the 5'-UTR shown in this study to be active in TR. The silencing effect of this 5'-UTR was shown to be independent of its position relative to its ORF, as transposition to a location 3' of the ORF did not affect TR. These results demonstrate for the first time in a unicellular organism that the 5' or the 3'-UTR of TR-associated transcripts play an important and conserved role in mediating TR in female gametocytes.