APPLICATION OF NOVEL VECTORS FOR GFP-TAGGING OF PROTEINS TO STUDY MICROTUBULE-ASSOCIATED PROTEINS

We describe the construction of pBact-NGFP and pBact-CGFP, two express ion vectors that incorporate green fluorescent protein (GFP) as a fluo rescent tag at the N- or C terminus of the produced protein. When tran sfected into recipient cells, GFP-tagged proteins can be visualised in the living cells using standard fluorescence microscopy techniques. U sing these expression vectors, we have produced GFP-tagged versions of the neuronal microtubule-associated proteins (MAP), MAP2c and Tau34, in a number of different cell types, Both GFP-MAP2c and GFP-Tau34 were fluorescent and retained their ability to bind to microtubules. The p Bact-NGFP and pBact-CGFP expression vectors represent a fast and conve nient way to produce fluorescently tagged polypeptides of selected seq uences encoding whole proteins or fragments for the analysis of functi on and dynamic events in living cells.