Functional expression and characterization of the mouse epitope tag-protein kinase C isoforms, α, βI, βII, γ, δ and ε

To simplify the detection of the exogenously produced protein kinase C (PKC) isoforms, we constructed the T7-Tag PKCα, βI, βII, γ, δ and ε expression plasmids. T7-Tag sequences (AlaSerMetThrGlyGlyGlnGlnMetGlyArg) were inserted at the 5′-end of the translational initiation site. Transient transfection of T7-Tag PKC expression plasmids into CV-1 cells increased the levels of PKC protein, and PKC activity. T7-Tag-PKCε, like native PKCε, transactivated the transcription of a NF-κB reporter construct. These results indicate that the plasmids encoding T7-Tag PKC are functional and may be useful to define the role PKC isoforms play in cell proliferation, differentiation and tumor promotion.