Suppose you have a 1-kb segment of cloned DNA that is suspecte...

Question

Suppose you have a 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 (1) determine if eukaryotic transcription factors (TF) bind to the fragment and, if so, (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 two items your supervisor has listed and to describe the kind of results that would indicate binding of TF to the DNA and the location of the binding.

Accepted Answer

Hey, everyone. Let's take a look at this question together. This essay is helpful in determining a few periodic transcription factors bind to the DNA fragment. Is it answer choice. A electrophrenic mobility shift assay. Answer choice B liras chain reaction, answer choice C restriction enzymes or answer choice D DNA recombination. Let's work this problem out together to try to figure out which of the following answer choices can be helpful in determining if E Caro transcription factors bind to the DNA fragment. So we know that the term assay is, is a test that is used to determine the amount of something. And in this case, we want to know if the eukaryotic transcription factors bind to the DNA fragment. So looking at our answer choices, we can eliminate some of them immediately since they are used for something other than determining if eukaryotic transcription factors bind to the DNA fragment. And we can eliminate answer choice B polymerase chain reaction. Since we note that Mary's chain reaction is a tool that is used for amplifying copies of DNA segments and cannot be used to determine if the transcription factors bind. And we can also eliminate answer choice C restriction enzyme. Since we know that restriction enzymes are enzymes that are used for cleaving DNA sequences or something such as DNA recombination. So answer choice C is incorrect. And lastly, we can eliminate answer choice D since we know that DNA recombination is a method that is used for combining DNA from multiple sources. And is not something that can be helpful in determining if eucaryotic transcription factors bind to the DNA fragment, which leaves us with answer choice. A as the only remaining answer choice as well as the correct answer. Since electrophrenic mobility shift assay can be helpful in determining if E cario transcription factors bind to the DNA fragment. Since the process of electrolytic mobility shift assay involves the use of a sample of DNA that contains nuclear extracts containing transcription factors. And then that sample of DNA is subject to gel electrophoresis, which we know is a process that is used to separate DNA mixtures by size. And in the case of our DNA sample, if those transcription factors are present and bound to the DNA, the DNA will migrate slowly in the gel electrophoresis. Since gel electrophoresis is a method that separates those DNA mixtures by size and the bound DNA meaning the DNA that contains those transcription factors is larger because of the transcription factors. So it will migrate slowly making answer choice. A electrolytic mobility shift assay. An essay that is helpful in determining if you carry out a transcription factors bind to the DNA fragment. I hope you found this video to be helpful. Thank you and goodbye.

Key Concepts

Eukaryotic Promoters

Transcription Factors (TFs)

Electrophoretic Mobility Shift Assay (EMSA)

Watch next