A virus with a circular double-stranded DNA chromosome contains approximately 10,000 bp. You want to begin characterizing this chromosome by making a map of the cleavage sites of three restriction endonucleases: EcoRI, Hind III, and BamHI. You digest the viral DNA under conditions that allow the endonuclease reactions to go to completion and then subject the digested DNA to electrophoresis on agarose to determine the lengths of the restriction fragments produced in each reaction. Based on the resulting data, draw a map of the viral chromosome indicating the relative positions of the cleavage sites for these restriction endonucleases: Endonuclease EcoRI HindIII BamHI EcoRI+HindIII EcoRI + BamHI HindIII+BamHI EcoRI + HindIII + BamHI Length of fragments (kb) 6.9,3.1 5.1, 4.4, 0.5 10.0 3.6, 3.3, 1.5, 1.1, 0.5 5.1, 3.1, 1.8 4.4, 3.3, 1.8, 0.5 3.3, 1.8, 1.5, 1.1, 0.5

A virus with a circular double-stranded DNA chromosome contains approximately 10,000 bp. You want to begin characterizing this chromosome by making a map of the cleavage sites of three restriction endonucleases: EcoRI, Hind III, and BamHI. You digest the viral DNA under conditions that allow the endonuclease reactions to go to completion and then subject the digested DNA to electrophoresis on agarose to determine the lengths of the restriction fragments produced in each reaction. Based on the resulting data, draw a map of the viral chromosome indicating the relative positions of the cleavage sites for these restriction endonucleases: Endonuclease EcoRI HindIII BamHI EcoRI+HindIII EcoRI + BamHI HindIII+BamHI EcoRI + HindIII + BamHI Length of fragments (kb) 6.9,3.1 5.1, 4.4, 0.5 10.0 3.6, 3.3, 1.5, 1.1, 0.5 5.1, 3.1, 1.8 4.4, 3.3, 1.8, 0.5 3.3, 1.8, 1.5, 1.1, 0.5

Biology Today and Tomorrow without Physiology (MindTap Course List)

5th Edition

ISBN:9781305117396

Author:Cecie Starr, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Christine Evers, Lisa Starr

Chapter10: Biotechnology

Section: Chapter Questions

Problem 1CT

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Restriction endonucleases are bacterial enzymes that cleave duplex (double-stranded) DNA at specific nucleotide sequences. The mode of replication of the animal virus SV40 has been investigated by using restriction endonucleases that cleave SV40 DNA into a number of unique segments. Like most viruses, SV40 DNA is circular. The map positions of the 11 fragments produced by a pair of restriction endonucleases are shown on the next page. Immediately following a 5 or 10 minute pulse of radioactively labeled thymidine, labeled SV40 molecules that have completed replication during the pulse are isolated. These newly replicated DNA molecules are digested by the restriction endonucleases and the resulting fragments are analyzed for the relative amounts of pulse label they contain. The results are in the table below. Assume that at the time the label was added there was a random population of replicating SV40 DNA molecules in all possible stages of synthesis. From the information given below,…
Restriction sites are palindromic; that is, they read the same in the5' to 3' direction on each strand of DNA. What is the advantage ofhaving restriction sites organized this way?
A plasmid DNA and a linear DNA (both of the same size) have one site for a restriction endonuclease. When cut and separated on agarose gel electrophoresis, plasmid shows one DNA band while linear DNA shows two fragments. Explain.
E 1 kb 1 kb DAK 6 kb Gene X E1 kb B 5 kb. Gene Z E₁kb 1 kb The figure above shows a linear chronosome containing Gene X and Gene Z and the location of sites for the restriction enzymes EcoRI (E) and BamHI (B). Refer to this map as you answer the question below. If you digest this DNA with EcoRI, then perform agarose gel electrophoresis, how many bands, and what sizes, would you expect to see? a) Two bands on the gel corresponding to the sizes 2 kb and 6 kb b) Three bands on the gel corresponding to the sizes 1 kb, 6 kb and 8 kb. c) Three bands on the gel corresponding to the sizes 1 kb, 5 kb and 6 kb d) Four bands on the gel corresponding to the sizes 1 kb, 2 kb, 5 kb and 6 kb e) Four bands on the gel corresponding to the sizes 2 kb, 3 kb, 4 kb and 8 kb
The restriction endonuclease NotI recognizes the octanucleotide sequence GCGGCCGC. Calculate the expected number of NotI cleavage sites in the bacteriophage l genome, a linear DNA duplex 48.5 kbp in length with a (G + C) content of 50%.
To estimate the number of cleavage sites in a particular piece of DNA with a known size, you can apply the formula N/4n where N is the number of base pairs in the target DNA and n is the number of bases in the recognition sequence of the restriction enzyme. If the recognition sequence for BamHI is GGATCC and the l phage DNA contains approximately 48,500 bp, how many cleavage sites would you expect?
The restriction endonuclease NotI recognizes the octanucleotide sequence GCGGCCGC. Calculate the expected number of NotI cleavage sites in the bacteriophage λgenome, a linear DNA duplex 48.5 kbp in length with a (G + C) content of 50%.
Assume that a circular plasmid is 3200 base pairs in length and has restriction sites for HindIII restriction enzyme at the following locations: 400, 700, 1400, 2600. Give the expected sizes of the restriction fragments following complete digestion.
You are studying a new plasmid, and you digest the plasmid with three restriction enzymes: Eco RI (E), Hindlll (H), and Xbal (X). You digest the plasmid DNA with each of the following combinations of enzymes and observe the results on an agarose gel. You are provided a partial plasmid map as shown below to the right. +18 a. What is the size of this plasmid in base pairs? b. What is the distance in base pairs between E1 and H? c. What is the distance in base pairs between E1 and X? d. What is the distance in base pairs between E2 and H? e. What is the distance in base pairs between E2 and X?
A 2.0kb bacterial plasmid ‘BS1030’ is digested with the restriction endonuclease Sau3A; the plasmid map is depicted in the diagram below and the Sau3A (S) restriction sites are indicated. Which of the following DNA fragments do you expect to see on an agarose gel when you run Sau3A-digested plasmid ‘BS1030’ DNA? a. 250 bp, 450 bp, 550 bp, 1.1 kb, 1.5 kb and 2.0 kb b. 2.0kb c. 250 bp, 400 bp, 450 bp, 500 bp and 550 bp d. 100 bp, 200 bp, 250 bp, 400 bp, 500 bp and 550 bp
When the restriction endonuclease EcoRI is used to digest a 10 kb DNA fragment, it produces 4 kb and 6 kb-sized fragments. Digesting the 10 kb fragment with BamHI yields three fragments, each ranging in size from one to three and a half kilobytes. Four pieces of 0.5, 1, 3 and 5.5 kb are formed after using both enzymes. Create a restriction map for this 10 kb piece of DNA using the information you have collected. Make a note of where the two enzymes cut, as well as the distances between the enzymes.
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