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New Fusion Trans-Activator Plasmids for Studying Signal Transduction,,,Pathways

ing systems include unique fusion trans-activator plasmids, a reporter plasmid that encodes the firefly luciferase gene and positive control plasmids. The fusion trans-activator plasmids express pathway-specific trans-activator proteins that consist of the yeast GAL4 DNA binding domain and the activation domains of various transcription factors. When a fusion trans-activator plasmid, reporter plasmid and uncharacterized gene are cotransfected into mammalian cells, either direct or indirect phosphorylation of the fusion trans-activator protein by the uncharacterized gene product will activate transcription of the luciferase gene from the reporter plasmid (figure 1).

To increase the utility of the PathDetect systems, Stratagene has designed the pFA-CMV plasmid for convenient insertion of the activation domain of any transcription factor fused to the yeast GAL4 DNA binding domain. The pFA-CMV plasmid was used to construct two new PathDetect fusion trans-activator plasmids, pFA-cFos and pFA-ATF2 plasmids, for studying certain signal transduction pathways converging at the transcription factors c-Fos and ATF2. These plasmids will expand our capacity to study various aspects of cellular signal transduction pathways. Each plasmid is highly purified, tested and available separately for use with the PathDetect systems.

Design and Testing of the pFA-CMV Plasmid

figure 2

Researchers can use the pFA-CMV plasmid (figure 2) for easy cloning of the activation domain of any transcription factor sequence and subsequent expression of GAL4 fusion proteins in a variety of mammalian cells. The pFA-CMV plasmid features (1) a multiple cloning site (MCS) that offers 10 unique, conveniently arranged restric
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