Rapid expression of polymorphic ovine prion proteins and studies on their protease sensitivity

We have used coupled and uncoupled in vitro transcription/translation to ex press rapidly aglycosyl ovine prion proteins from ovine genomic DNA genotyp ed for scrapie susceptible and nonsusceptible polymorphisms. Unlike previou s in vitro studies of prion proteins, this method does not require cloning or laborious extractions [2]. To our knowledge, this is the first report of ovine PrP expression at low (ng) levels under the control of an Escherichi a coil promoter and ribosome binding site both coded for in the polymerase chain reaction primer. The rapidity of this approach could form the basis o f a high throughput screening assay for PrP interactions, as proteins were expressed in a matter of hours from genomic DNA as the starting material. T here was no difference observed in proteinase K sensitivity between prion t ranslation products containing either scrapie susceptible or nonsusceptible polymorphisms. (C) 1999 Elsevier Science Inc.