Tg(Prnp)940Zbz

Mice with this transgene express a Myc epitope tagged prion protein under control of the regulatory sequences of the mouse prion protein gene (Prnp) contained in the "half-genomic" vector phgPrp, which includes 6 kb of 5'- and 2.2 kb of 3'-flanking DNA, exon 1, intron 1, and exon 2 joined to exon 3 of the mouse Prnp gene. The expressed protein is a mouse prion protein with a sub-C terminal human Myc epitope (EQKLISEEDL) following Ser230, just before the C terminal membrane anchor of the PRNP precursor protein that in the mature protein is replaced by a GPI anchor; the Myc epitope is followed by a repetition of the five amino acid sequence culminating in Ser230. By Southern blot analysis it was estimated that mice of this transgenic line carry six copies of the transgene per haploid genome. Transcription of the transgene was confirmed by northern blot analysis of total brain RNA. Transgenic mice with a Prnp null background were shown by western blot analysis of whole tissue extracts to express ~1.6-fold the wild-type level of PRNP in brain and about 0.5-fold the wild-type level in spleen. Appropriate localization of the Myc-tagged protein to detergent resistant membranes was demonstrated by its presence, detected by western blot analysis, in the same centrifugal gradient fractions of cerebellar DRM preparations as flotillin 2. (J:32214, J:183308)