Chapter 8, Problem 27P

Suppose you have a $\text{1-kb}$ segment of cloned DNA that is suspected to contain a eukaryotic promoter including a TATA box, a CAAT box, and an upstream GC-rich sequence. The clone also contains a gene whose transcript is readily detectable. Your laboratory supervisor asks you to outline an experiment that will ( $\text{1}$) determine if eukaryotic transcription factors (TF) bind to the fragment and, if so, ( $\text{2}$) identify where on the fragment the transcription factors bind. All necessary reagents, equipment, and experimental know-how are available in the laboratory. Your assignment is to propose techniques to be used to address the three items your supervisor has listed and to describe the kind of results that would indicate binding of TF to the DNA, the location of the binding. (Hint: The techniques and general results are discussed in this chapter.)