Tg(Pcna*K164R)1Mdsc

A genomic fragment was generated by SphI digestion that contained about 3 kb of the 5' promoter region of the murine Pcna gene, all of its exon and intron sequences, and 875 bp of its 3'untranslated region. An A to G mutation was introduced by site-directed mutagenesis to change codon 164 from lysine to arginine (AGG) and to generate a new EcoNI restriction site. Real-time PCR on B cell extracts showed an approximately two- to eight- fold increase of steady-state mRNA compared to mRNA levels derived from the endogenous Pcna locus. Immunoblot assays on B cell extracts confirmed higher expression of the transgene. The point mutation is predicted to interfere with ubiquitylation of the protein product. (J:141098)