Genetic Analysis: An Integrated Approach (3rd Edition)
Genetic Analysis: An Integrated Approach (3rd Edition)
3rd Edition
ISBN: 9780134605173
Author: Mark F. Sanders, John L. Bowman
Publisher: PEARSON
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Chapter 8, Problem 26P

DNA footprint protection (described in Research Technique 8 .1 ) is a method that determines whether proteins bind to a specific sample of DNA and thus protect part of the DNA from random enzymatic cleavage by DNase I. A 400-bp segment of cloned DNA is thought to contain a promoter. The cloned DNA is analyzed by DNA foot printing to determine if it has the capacity to act as a promoter sequence. The gel shown below has two lanes, each containing the cloned 400-bp DNA fragment treated with DNase I to randomly cleave unprotected DNA. Lane 1 is cloned DNA that was mixed with RNA polymerase II and several TFII transcription factors before exposure toDNase I. Lane 2 contains cloned DNA that was exposed only to DNase I. RNA pol II and TFIIs were not mixed with DNA before adding DNase I.

a. Explain why this gel provides evidence that the cloned DNA may act as a promoter sequence.

b. Approximately what length is the DNA region protected by RNA pol II and TFIIs?

c. What additional genetic experiments would you suggest to verify that this region of cloned DNA contains a functional promoter?

Chapter 8, Problem 26P, DNA footprint protection (described in Research Technique 8.1) is a method that determines whether

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There may be more than one correct answer for each Which of the following motifs can be found in DNA-binding proteins?  a.) helix-loop-helix b.) zinc fingers c.) leucine zippers d.) CpG islands   Which of the following motifs can be used to bind protein dimers?  a.) helix-loop-helix b.) zinc fingers c.) leucine zippers d.) CpG islands   Which of the following are correct regarding enhancers?  a.) Enhancers may be located a great distance from the initiation site they regulate. b.) Enhancers can be either upstream or downstream from the initiation site. c.) Enhancers directly bind DNA and prevent binding of activators to the DNA. d.) Enhancers may function in either orientation (forward or backward).
When a region of DNA that contains the genetic information for a protein is isolated from a bacterial cell and inserted into a eukaryotic cell in a proper position between a promoter and a terminator, the resulting cell usually produces the correct protein. But when the experiment is done in the reverse direction (eukaryotic DNA into a bacterial cell), the correct protein is often not produced. Can you suggest an explanation?
Suppose you have a 1-kb segment of cloned DNA that is suspected to contain a eukaryotic promoter including a TATA box, a CAT 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. (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. Complete the outline of an experiment, which determines if eukaryotic transcription factors (TF) bind to the fragment. Assume all necessary reagents, equipment, and experimental how are available in the laboratory. Drag the terms on the left to the appropriate blanks on the right to complete the sentences. Not all terms will be used. For the band shift assay, two samples of the DNA fragment are analyzed by agarose gel electrophoresis: one sample is…

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Genetic Analysis: An Integrated Approach (3rd Edition)

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