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Functional expression and characterization of the mouse epitope tag-protein kinase C isoforms, alpha, beta I, beta
1Department of Human Oncology, Medical School, University of Wisconsin-Madison 53792, USA. ctseng@students.wisc.edu
Abstract:
To simplify the detection of the exogenously produced protein kinase C (PKC) isoforms, we constructed the T7-Tag PKC alpha, betaI, betaII, gamma, delta and epsilon 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-PKC epsilon, like native PKC epsilon, transactivated the transcription of a NF-kappa B 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.