To evaluate the fruit fly as a model for studying neurodegenerative diseases caused by prions, transgenic flies were generated by introducing the Syrian hamster prion protein (SHaPrP) gene into the Drosophila melanogaster germ line by P element-mediated transformation. Nine transgenic lines were isolated; induction of transgenes that had been placed under the control of the Drosophila heat shock promoter, hsp 70, resulted in the synthesis of full-length SHaPrP. The relative molecular weight of the recombinant protein was lower than that of authentic SHaPrP due to incomplete processing of Asn-linked CHOs. To determine the cellular localization of SHaPrP, Drosophila Schneider line 2 cells were transfected with the same constructs used for fly transformation. Heat shock induced SHaPrP was anchored to the surface of S2 cells by a glycolipid, demonstrating that the carboxy-terminal glycolipidation signal of SHaPrP is recognized by this evolutionarily distant host. When SHaPrP was synthesized in transgenic flies constitutively by subjecting them to heat pulses continuously, no difference in their lifespans compared with controls was detected. Furthermore, expression of SHaPrP for 20 days did not produce protease resistant SHaPrP, which is the major and possibly only component of the infectious prion. In contrast to transgenic mice overexpressing SHaPrP, which develop a profound neuromyopathy, no disease phenotype was associated with expression of SHaPrP over the entire lifespan of transgenic flies.