Biology: Life on Earth with Physiology (11th Edition)
11th Edition
ISBN: 9780133923001
Author: Gerald Audesirk, Teresa Audesirk, Bruce E. Byers
Publisher: PEARSON
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Chapter 14.4, Problem 1TC
Restriction enzymes are isolated from bacteria. Why would bacteria synthesize enzymes that cut up DNA? (Hint: Bacteria can be infected by viruses called bacteriophages; see Chapter 12.) Why wouldn't a bacterium's restriction enzymes destroy the DNA of its own chromosome?
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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?
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 endonuclease and ligase are two types
of enzymes used in the process of genetic
engineering, i.e., the manipulation of genes. The
restriction endonuclease differs from ligase in that it
breaks the DNA at ends, while ligase causes
the breaks in DNA from interior
joins the fragments of DNA, while ligase
breaks the DNA into fragments
breaks the DNA at specific points, while the
ligase joins the fragments of DNA
breaks the DNA apart at each nucleotide,
while ligase use the pieces to translate
Chapter 14 Solutions
Biology: Life on Earth with Physiology (11th Edition)
Ch. 14.1 - define biotechnology?Ch. 14.1 - Prob. 2CYLCh. 14.2 - describe natural processes that recombine DNA,...Ch. 14.3 - Guilty or Innocent? When biological evidence was...Ch. 14.3 - For any single person, a given STR always has...Ch. 14.3 - Prob. 2CSCCh. 14.3 - There are many other applications in which DNA...Ch. 14.3 - Prob. 1CYLCh. 14.3 - Prob. 2CYLCh. 14.3 - Prob. 3CYL
Ch. 14.4 - Restriction enzymes are isolated from bacteria....Ch. 14.4 - explain how genes are inserted into a plasmid, and...Ch. 14.4 - Prob. 2CYLCh. 14.5 - Prob. 1HYEWCh. 14.5 - describe the advantages of genetically modified...Ch. 14.5 - list some examples of how GM animals might be...Ch. 14.5 - Prob. 3CYLCh. 14.6 - Prob. 1CYLCh. 14.6 - explain how knowledge of the genomes of humans and...Ch. 14.7 - Prob. 1TCCh. 14.7 - explain how biotechnology is used to diagnose both...Ch. 14.7 - describe the procedures and advantages of gene...Ch. 14.8 - Genetic engineering is used both in food crops and...Ch. 14.8 - explain why people might be opposed to the use of...Ch. 14.8 - envision circumstances in which it would be...Ch. 14.8 - Prob. 1CSRCh. 14.8 - Prob. 2CTCh. 14 - Prob. 1MCCh. 14 - Prob. 2MCCh. 14 - Prob. 3MCCh. 14 - A restriction enzyme a. cuts DNA at a specific...Ch. 14 - Prob. 5MCCh. 14 - Prob. 1FIBCh. 14 - _________is the process whereby bacteria pick up...Ch. 14 - The _______ is a technique tor multiplying DNA in...Ch. 14 - Matching DNA samples in forensics uses a specific...Ch. 14 - Prob. 5FIBCh. 14 - Prob. 1RQCh. 14 - Prob. 2RQCh. 14 - Prob. 3RQCh. 14 - Prob. 4RQCh. 14 - Prob. 5RQCh. 14 - How does gel electrophoresis separate pieces of...Ch. 14 - Prob. 7RQCh. 14 - Prob. 8RQCh. 14 - Prob. 9RQCh. 14 - Prob. 10RQCh. 14 - As you may know, many Insects have evolved...Ch. 14 - Prob. 2AC
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- An important feature of restriction enzymes is that each enzyme only recognizes a specific palindrome and cuts the DNA only at that specific sequence of bases. A palindromic sequence can be repeated a number of times on a strand of DNA, and the specific restriction enzyme will cut all those palindromes, no matter what species the DNA comes from. A linear DNA molecule is represented below. The DNA is represented by one line, although in actuality, DNA has two strands. If the DNA molecule has two restriction sites, specifically two repeats of a specific palindrome sequence, A and B, for a specific restriction enzyme: How many fragments would be produced if the DNA is cut by that enzyme? Number each fragment Which fragment would be the largest? Which fragment would be the smallest?arrow_forwardA restriction map lists the locations of DNA sequences that are cut by a particular restriction enzyme for a piece of DNA, such as a chromosome or a plasmid. Restriction maps are important when generating a construct for experimental use. Digesting the DNA sequence with the restriction enzymes will result in fragmented DNA of predictable sizes, based on the restriction map, that allow a researcher to analyze if his or her construct was generated correctly when visualized using gel electrophoresis. Use the linear restriction map to predict where bands would be expected on a gel if a digest is performed using the specified restriction enzymes. Assume that there is enough restriction enzyme that every possible restriction site on each molecule of DNA will be cut.arrow_forwardIf restriction endonucleases are produced by bacteria within a host, why don’t these enzymes chew up the genomic DNA of their host? What is the role of DNA methyltransferase in this?arrow_forward
- You have a recombinant plasmid containing a vector and a segment of foreign DNA, both equal sizes. Draw a picture of this recombinant plasmid labeling foreign and vector regions. Where the foreign DNA meets the vector, there is a cut site for restriction enzyme ABC1. When the recombinant plasmid is cut by ABC1, how many fragments do you expect to be produced? Identify these fragments.arrow_forwardYou isolated a 10,500 base pair plasmid (supercoiled, cccDNA) from E. coli. The plasmid contains one unique recognition site for EcoRI, a restriction enzyme. Restriction enzymes recognize a specific sequence and cut both strands of the DNA at that sequence (Chapter 7). Brief DNase I treatment breaks one (or very few) phosphodiesterbonds in each DNA molecule, leaving the double-stranded DNA with one strand broken but the other strand intact, i.e “nicked.” You briefly incubated the cccDNA at 37°C in four separate reactions containing the components listed below. You ran each reaction sample on an agarose gel and visualized the DNA using ethidium bromide and UV light. The reactions included the appropriate buffer and ATP when required. An agarose gel containing four lanes of possible products is given below. Please refer to Figs. 4-26 and 4-27 in Watson for reference. For each reaction, indicate which lane of the gel contains the products that you would expect to see on your…arrow_forwardRestriction enzymes in bacterial cytoplasm cut injected bacteriophage DNA wherever certain sequences occur. Why do you think these enzymes do not chop up the bacterial chromosome, which is exposed to the enzymes in the cytoplasm?arrow_forward
- In the formation of recombinant DNA, a restriction endonuclease cuts a bacterial plasmid to give sticky ends. The DNA segments that are to be added to the plasmid are cleaved with the same restriction endonuclease. What aresticky ends and why is it important that the target DNA and the plasmid it will be incorporated into have complementary sticky ends?arrow_forwardA small DNA molecule was cleaved with several different restriction nucleases, and the size of each fragment was determined by gel electrophoresis.The following data were obtained. (a) Is the original molecule linear or circular?(b) Draw a map of restriction sites (showing distances between sites) that isconsistent with the data given.(c) How many additional maps are compatible with the data?(d) What would have to be done to locate the cleavage sites unambiguouslywith respect to each other?arrow_forwardA 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.arrow_forward
- Now that you’ve isolated the gene and made lots of copies, you need to insert the gene into something you can manipulate and move into your lab strain of E. coli. You have access to the following vectors, either pBR322 or pUC19 (look them up here to see a map https://www.neb.com/products/dna-plasmids-and-substrates Please note the restriction sites in BOLD appear only once in the plasmid). You may use either vector. What is a vector? Which vector did you choose? Explain why you made that choice. What enzyme(s) will you use to place your fragment into the vector? Explain why you chose these enzymes. How will you move this construct into coli? What phenotype will tell you if the coli have taken up the plasmid? What phenotype will tell you if the coli have taken up the plasmid with the gasP gene?arrow_forwardIn Cohen-Boyer’s recombinant DNA procedure, ___i___ must be used for both the bacterial DNA and the amphibian DNA ___ii___ a) the same restriction enzyme; so that the restriction sites are identical in the DNA of each species b) different restriction enzymes; So that the genes outside the restriction site are maintained c) different restriction enzymes; to ensure that the newly introduced genes are maintained in the bacterial DNA d) the same restriction enzyme; to ensure that the newly formed DNA can replicatearrow_forwardIf a restriction enzyme cuts at this DNA sequence: TACGGAT, in general what is this sequence called?arrow_forward
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