( on factors are not able to bind efficie...)In Vivo Signal Transduction Pathway Reporting Systems

on factors are not able to bind efficiently to the yeast-derived GAL4 UAS sequence. The high specificity and low background of the PathDetect systems have additional applications, including mapping a protein's position in the signal transduction pathway by introducing known inhibitors of the pathway and screening for drugs that promote or inhibit the activity of JNK, MAPK, PKA or other upstream signaling molecules (such as Ras, raf and JNKK).

Conclusions

The PathDetect in vivo signal transduction pathway reporting systems can be used to determine and map the signal transduction pathway for a gene product and to study the effects of extracellular stimuli on the JNK, MAPK or PKA signal transduction pathways. Researchers can use these systems to evaluate the in vivo role of a new gene product by the activation, inhibition or regulation of the selected signal transduction pathway. The firefly luciferase gene is used as the reporter gene in the PathDetect reporter plasmid. Therefore, the activation of these signaling pathways can be readily measured by the luciferase assay, which is convenient, sensitive and quantitative. The kits include fusion transactivator plasmids, reporter plasmids and positive and negative control plasmids that are highly purified and transfection ready.

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