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Ch. 10 - Eukaryotic Chromosome Abnormalities and Molecular Organization
Sanders - Genetic Analysis: An Integrated Approach 3rd Edition
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172Not the one you use?Change textbook
Chapter 10, Problem 27b

Genomic DNA from the nematode worm Caenorhabditis elegans is organized by nucleosomes in the manner typical of eukaryotic genomes, with 145 bp encircling each nucleosome and approximately 55 bp in linker DNA. When C. elegans chromatin is carefully isolated, stripped of nonhistone proteins, and placed in an appropriate buffer, the chromatin decondenses to the 10-nm fiber structure. Suppose researchers mix a sample of 10-nm–fiber chromatin with a large amount of the enzyme DNase I that randomly cleaves DNA in regions not protected by bound protein. Next, they remove the nucleosomes, separate the DNA fragments by gel electrophoresis, and stain all the DNA fragments in the gel.
Explain the origin of DNA fragments seen in the gel.

Verified step by step guidance
1
Step 1: Understand the structure of chromatin in Caenorhabditis elegans. Chromatin consists of nucleosomes, which are DNA segments of 145 base pairs (bp) wrapped around histone proteins, and linker DNA, which is approximately 55 bp long and connects adjacent nucleosomes. The nucleosomes protect the DNA they encircle from enzymatic cleavage.
Step 2: Recognize the role of DNase I in the experiment. DNase I is an enzyme that randomly cleaves DNA in regions not protected by bound proteins, such as the linker DNA between nucleosomes. The DNA wrapped around histones is shielded from DNase I activity.
Step 3: Consider the experimental setup. After treating the chromatin with DNase I, the researchers remove the nucleosomes, leaving behind DNA fragments that were protected by histones and fragments that were cleaved in the linker regions.
Step 4: Analyze the DNA fragments using gel electrophoresis. Gel electrophoresis separates DNA fragments based on their size. The protected DNA fragments (145 bp) and cleaved linker DNA fragments (approximately 55 bp or smaller) will appear as distinct bands on the gel.
Step 5: Explain the origin of the DNA fragments seen in the gel. The bands correspond to DNA fragments protected by nucleosomes (145 bp) and smaller fragments resulting from DNase I cleavage of linker DNA. The pattern reflects the periodic organization of chromatin into nucleosomes and linker regions.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Nucleosome Structure

Nucleosomes are the fundamental units of chromatin, consisting of DNA wrapped around a core of histone proteins. Each nucleosome typically contains about 145 base pairs of DNA, which is protected from enzymatic cleavage. Understanding nucleosome structure is crucial for explaining how certain regions of DNA remain intact during experiments involving DNase I, as the nucleosomes shield the DNA from being cut.
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DNase I Activity

DNase I is an enzyme that cleaves DNA at sites that are not protected by proteins, such as histones in nucleosomes. When chromatin is treated with DNase I, the enzyme will cut the linker DNA and any exposed regions of DNA, leading to the generation of fragments. This concept is essential for understanding the resulting DNA fragments observed in gel electrophoresis after nucleosome removal.
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Gel Electrophoresis

Gel electrophoresis is a laboratory technique used to separate DNA fragments based on their size. When DNA fragments are subjected to an electric field in a gel matrix, smaller fragments migrate faster than larger ones. This method allows researchers to visualize the DNA fragments generated by DNase I cleavage, providing insights into the organization and accessibility of chromatin in C. elegans.
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Related Practice
Textbook Question

DNase I cuts DNA that is not protected by bound proteins but is unable to cut DNA that is complexed with proteins. Human DNA is isolated, stripped of its nonhistone proteins, and mixed with DNase I. Samples are removed after 30 minutes, 1 hour, and 4 hours and run separately in gel electrophoresis. The resulting gel is stained to make all DNA fragments in it visible, and the results are shown in the figure. DNA fragment sizes in base pairs (bp) are estimated by the scale to the left of the gel. Examine the gel results and speculate why longer DNase I treatment produces different results.

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Textbook Question

DNase I cuts DNA that is not protected by bound proteins but is unable to cut DNA that is complexed with proteins. Human DNA is isolated, stripped of its nonhistone proteins, and mixed with DNase I. Samples are removed after 30 minutes, 1 hour, and 4 hours and run separately in gel electrophoresis. The resulting gel is stained to make all DNA fragments in it visible, and the results are shown in the figure. DNA fragment sizes in base pairs (bp) are estimated by the scale to the left of the gel. Draw a conclusion about the organization of chromatin in the human genome from this gel.

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Textbook Question

Genomic DNA from the nematode worm Caenorhabditis elegans is organized by nucleosomes in the manner typical of eukaryotic genomes, with 145 bp encircling each nucleosome and approximately 55 bp in linker DNA. When C. elegans chromatin is carefully isolated, stripped of nonhistone proteins, and placed in an appropriate buffer, the chromatin decondenses to the 10-nm fiber structure. Suppose researchers mix a sample of 10-nm–fiber chromatin with a large amount of the enzyme DNase I that randomly cleaves DNA in regions not protected by bound protein. Next, they remove the nucleosomes, separate the DNA fragments by gel electrophoresis, and stain all the DNA fragments in the gel.

Approximately what range of DNA fragment sizes do you expect to see in the stained electrophoresis gel? How many bands will be visible on the gel?

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Textbook Question

Genomic DNA from the nematode worm Caenorhabditis elegans is organized by nucleosomes in the manner typical of eukaryotic genomes, with 145 bp encircling each nucleosome and approximately 55 bp in linker DNA. When C. elegans chromatin is carefully isolated, stripped of nonhistone proteins, and placed in an appropriate buffer, the chromatin decondenses to the 10-nm fiber structure. Suppose researchers mix a sample of 10-nm–fiber chromatin with a large amount of the enzyme DNase I that randomly cleaves DNA in regions not protected by bound protein. Next, they remove the nucleosomes, separate the DNA fragments by gel electrophoresis, and stain all the DNA fragments in the gel.

How do the expected results support the 10-nm–fiber model of chromatin?

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Textbook Question

A small population of deer living on an isolated island is separated for many generations from a mainland deer population. The populations retain the same number of chromosomes but hybrids are infertile. One chromosome (shown here) has a different banding pattern in the island population than in the mainland population.

Describe how the banding pattern of the island population chromosome most likely evolved from the mainland chromosome. What term or terms describe the difference between these chromosomes?

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Textbook Question

A small population of deer living on an isolated island is separated for many generations from a mainland deer population. The populations retain the same number of chromosomes but hybrids are infertile. One chromosome (shown here) has a different banding pattern in the island population than in the mainland population.

Draw the synapsis of these homologs during prophase I in hybrids produced from the cross of mainland with island deer.

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