Functional expression and characterization of the mouse epitope tag-protein kinase C isoforms, alpha, beta I, beta II, gamma, delta and epsilon.
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.
Tseng
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Department of Human Oncology, Medical School, University of Wisconsin-Madison 53792, USA. ctseng@students.wisc.edu
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D000595:Amino Acid Sequence; D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D015870:Gene Expression / Q000235:genetics*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011493:Protein Kinase C / Q000235:genetics* / Q000378:metabolism; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D016324:Sequence Tagged Sites
D000939:Epitopes; D007527:Isoenzymes; D004247:DNA; C497259:Prkcd protein, mouse; C497256:Prkce protein, mouse; C087857:protein kinase C gamma; C496630:Prkca protein, mouse; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon