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Mammalian expression systems have been used extensively for recombinant rabies virus (RV) production. However, because of the severe limitations imposed by the requirement for adaptation of these systems to growth on mammalian cells and the resulting inability of the RV glycoproteins (G) to interact efficiently with the T-cell receptors of the mammalian host, a system capable of producing functional native G without the mutations commonly required in adapting the virus to growth on mammalian cells has yet to be developed. In this work, we demonstrate that the well-known yeast system, Pichia pastoris, can be used to express RV glycoprotein with cell types that can serve as useful substrates for a further amplification in cell culture. Native G and rabies virus-like particles (VLPs) were produced efficiently with glycosylation patterns as they occur on the surface of the native virus. The glycoprotein was expressed at high levels of the order of 100 mg/L of culture. Immunological assays indicate that the heterologous G displays the correct topography and natural conformational

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