Abstract: To construct a kind of recombinant plasmid with ovine prion protein gene （PRNP） and transfect the plasmid into C6 cell line in order to study the physiological function of normal prion protein and the pathogenesis in prion diseases. The gene sequence encoding ovine prion gene was amplified by PCR and cloned into eukaryotic expression vector pEGFP-N1 with EGFP reported gene encoding enhanced green florescence protein, and then identified by restriction enzyme. The recombinant plasmid pEGFP-PRNP was transfected into C6 cell line by using lipofectin method. We successfully construct the expression vector of recombinant plasmid pEGFP-N1 and it can be steadily expressed in form of fused protein in C6 cell line. It is suggested that the expression of recombinant plasmids pEGFP-PRNP in C6 cell line will provide a platform for researching expression of prion in cell and further contribute to study on prion diseases at cell level in future.