(4.2) In an in vitro transcription reaction a high concentration of Gal4 is used
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Question
(4.2) In an in vitro transcription reaction a high concentration of Gal4 is used to inhibit ("squelch") transcription from a promoter that is activated by the transcriptional activator Gcn4. You have isolated a yet unknown activity M that has anti-squelching activity, i.e. addition of M alleviates squelching of Gen4-activated transcription. Propose an in vitro transcription experiment that allows you to test the hypotheses that M is an inhibitor of Gal4 (H) 34 Gal 8 0, for instance 4 and not Mediator. Describe the expected outcomes of your experiment for both hypotheses 34 if H is true, and if M is Mediator.Explanation / Answer
To test whether M is an inhibitor, we could perform the following experiment:
Rate of reaction should be measured while performing the experiment by adding different quantities of M and the same should be recorded in a table. In other experiment, M should not be used in the experiment. Then, we will check if the addition of M has a significant impact on the rate of reaction. The hypothesis will be the following:
Ho: There is no significant impact on the rate of reaction with addition of M
H1: There is a significant impact on the rate of reaction with addition of M
Expected Outcomes if M is an inhibitor: There will be a decrease in the rate of reaction with addition of M. We could test it by perfoming a t-test. The p-value should come out to be <0.05.
If M is Mediator: There will not be any significant impact on the rate of reaction with addition of M. Null hypothesis will prevail.
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