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

Ras-responsive.11 The pFA-cFos plasmid was constructed by inserting the DNA fragment encoding residues 208 to 313 of rat c-Fos12 into the BamH I and EcoR I sites of pFA-CMV. Our results (figure 3A) in transient transfection assays demonstrate that the MEKK protein activates the GAL4-c-Fos fusion protein expressed from the pFA-cFos plasmid. The mechanism of this activation is not yet characterized. PKA, MEK1 and other extracellular stimuli failed to activate the GAL4-c-Fos fusion protein (data not shown).

figure 3B

ATF2, or activating transcription factor 2, belongs to a large family of transcription factors that recognize the ATF2/CRE elements identified in the adenovirus early gene promoters.8-10,13 ATF2 forms homodimers as well as heterodimers with other members of the ATF, Jun and Fos transcription factors. The regulation of ATF2 is not fully characterized. However, phosphorylation of Thr-69 and Thr-81 in ATF2 by JNK is known to activate the ATF2 transcription factor.13 In order to construct pFA-ATF2 (1-96), the DNA fragment encoding amino acid residues 1-96 of human ATF214,15 was inserted into the BamH I and EcoR I sites of the pFA-CMV vector. Consistent with previous reports, the GAL4-ATF2 fusion protein expressed from the pFA-ATF2 plasmid was strongly activated by the MEKK protein in transient transfection assays (figure 3A).

Conclusions

Stratagene has constructed three new fusion trans-activator plasmids for use with the PathDetect in vivo signal transduction pathway reporting systems. All three of these plasmids can be used in a variety of mammalian cell lin
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Related biology technology :

1. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
2. Stable HeLa Luciferase Reporter Cell Lines Expressing GAL4 Fusion Transactivators
3. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
4. Comparison of Performance between the AlphaQuest - HTS and the Fusion α Multilabel Reader for Detection of cAMP
5. Mobile Phase Optimization for the Analysis of an Antibody-Based Fusion Protein and Aggregates using a TSKgel Super SW3000 Size-Exclusion Column
6. New Reporter Plasmids for Studying Interferon-Stimulated Signal Transduction Pathways
7. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
8. Optimizing Transfection Conditions for Studying Signal Transduction Pathways
9. Enzyme Immunoassay for Studying Intracellular Levels of cAMP
10. Assess the In Vivo Activation of Signal Transduction Pathways with PathDetect Reporting Systems
11. Signal Transduction Reporting Systems Using Cis-Acting Enhancer Elements
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