Anticipated Results & Significance

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Results and their significance for understanding the mechanism of immune response

Transfected cells have now TLR receptor (4 or 5) and our construct under NFκB promoter. We expect that after stimulation (with LPS or flagellin) cytoplasmic NFκB become active, migrate into nucleus and induce expression of our construct. Accumulating of dominant negative protein should block downstream signalling pathway. Concentration of active transcription factor should decrease and no more product should be synthesized. The rate of transcription and translation should be detected with detection systems - decrease of luminescence, free NFκB and::: (depends on detection system). In case that our construct is followed by PEST (degradation) sequence, it is expected that product should degrade. If stimulus is still present, NFκB will become active. Our product will be synthesized again.


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