Tg(Pcp2-PKCI)1Cidz

A minigene containing a PKC inhibitor was made by hybridization of two complementary synthetic oligonucleotides, each 50 bases in length, producing a double-stranded product with overhangs compatible with BamHI restriction sites. The minigene, is expressed under the control of a Purkinje cell specific promoter, and encodes a 13 amino acid peptide (not including the start methionine) corresponding in sequence to PKC[19-31] slightly truncated version of the pseudosubstrate domain of PKC, PKC[19-36] This version has been demonstrated in vitro to be a more potent inhibitor of PKC (by 60%) than the longer version, but both were shown by biochemical assay to block selectively the complete range of isotypes present in PKC purified from whole brain. Expression of the inhibitory peptide in Purkinje cells was confirmed by an electrophysiological assay of a phorbol ester-dependent effect that is attenuated in cerebellar cultures prepared from L7-PKCI mice. (J:108739)