Study Guide for Campbell Biology
11th Edition
ISBN: 9780134443775
Author: Lisa A. Urry, Michael L. Cain, Steven A. Wasserman, Peter V. Minorsky, Jane B. Reece, Martha R. Taylor, Michael A. Pollock
Publisher: PEARSON
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Textbook Question
Chapter 20, Problem 4IQ
- a. When PCR is used to prepare a DNA fragment for cloning, what sequence is often included in the primers?
- b. What type of gene is often included in the plasmids for gene cloning that helps identify the cells that have taken up a plasmid?
- c. Why even bother cloning genes in cells, given that PCR produces so many copies so fast?
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With the use of well-illustrated diagrams, reconstruct the entire cloning process by explaining different stages of the cloning process that involves the following:a. Isolation of target DNA fragments (often referred to as inserts)b. Ligation of inserts into the plasmid, creating recombinant molecules c. Transformation of recombinant plasmids into bacteria or other suitable host for propagationd. Screening/selection of hosts containing the intended recombinant plasmid. For this stage(d), discuss the importance of a second marker that can be used for screening of genomic DNA for colonies containing the pka-1 under the principle of insertional inactivation. This should be properly explained using all the attributes of the plasmid described above.
Describe how restriction enzymes like EcoR1 are used to create recombinant plasmids and what the process is for using these plasmids to replicate a piece of target DNA. Include information about how to create sticky ends, the makeup of the bacterial plasmid and how to tell if the gene was successfully inserted in the plasmid and if the plasmid has been transformed by the bacteria. You may use a drawing to enhance your description.
A.How could endonucleases interfere with the transformation procedure?
B. Does supercoiled or nicked plasmid get transformed more efficiently? Why?
Chapter 20 Solutions
Study Guide for Campbell Biology
Ch. 20 - In what ways would third-generation sequencing be...Ch. 20 - The following schematic diagram depicts an...Ch. 20 - Which of the following DNA sequences would most...Ch. 20 - a. When PCR is used to prepare a DNA fragment for...Ch. 20 - a. What are some of the benefits of determining...Ch. 20 - Prob. 6IQCh. 20 - What are some of the practical and ethical...Ch. 20 - Prob. 8IQCh. 20 - Prob. 1SYKCh. 20 - Fill in the table on the previous page on the...
Ch. 20 - Prob. 3SYKCh. 20 - Prob. 1TYKCh. 20 - Prob. 2TYKCh. 20 - Gel electrophoresis is a means of separating...Ch. 20 - Prob. 4TYKCh. 20 - Prob. 5TYKCh. 20 - The following segment of DNA has restriction sites...Ch. 20 - Prob. 7TYKCh. 20 - Prob. 8TYKCh. 20 - Prob. 9TYKCh. 20 - Prob. 10TYKCh. 20 - Prob. 11TYKCh. 20 - Which enzyme is used in the polymerase chain...Ch. 20 - Prob. 13TYKCh. 20 - STRs (short tandem repeats) are a valuable tool...Ch. 20 - Prob. 15TYKCh. 20 - Which of the following has the greatest potential...Ch. 20 - Prob. 17TYKCh. 20 - Petroleum-lysing bacteria are being engineered for...Ch. 20 - Prob. 19TYKCh. 20 - Prob. 20TYK
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- A. A plasmid is shown with the locations of various restriction enzyme sites labeled. If you cut the plasmid with Xhol and Xbal, which lane of the agarose gel represents the DNA fragments you would expect from the digestion? B. If you now decide to cut the plasmid with EcoRI, how many fragments will be produced and what will their sizes be? C. When running DNA samples on agarose gel, an electric field is applied. Towards which electrode will the DNA migrate and why?arrow_forwarda. Using nucleotide letters, show the kind of cut that could be made on a DNA molecule to circularize it into a plasmid. b. What are restriction length polymorphisms, and how are they used?arrow_forwarda. What determines the size (length) of the primary PCR product? b. What might a successful gel check of a PCR reaction look like?arrow_forward
- The temperature at which the primers and target DNA hybridize may be changed to influence the stringency of PCR amplification. What effect will changing the hybridization temperature have on the amplification? Let's say you have a certain yeast gene A and want to check whether it has a human equivalent. How might managing the hybridization's rigor benefit you?arrow_forwardA. Please briefly explain how Polymerase Chain Reaction works to amplify DNA. B. Please briefly explain what gel electrophoresis is and how it works to separate a mixed sample of macromolecules like DNA. C. Briefly describe what a plasmid is, and how it can be used to transform bacteria like E. coli.arrow_forwardMost PCR reactions do not use the more expensive types of DNA polymerase, which have DNA proofreading. How might this be a problem in accurately copying specific DNA sequences of the target gene?arrow_forward
- Primers can sometimes bind and target the wrong gene, especially if the primers are allowed to bind to the DNA strands at a low temperature. PCR also preferentially amplify short segments of DNA. Would it be important to actually run the cDNA after the PCR on a DNA gel in order to check for a PCR product of the predicted size for the insulin gene? Why or why not? How many more genes in the plasmid (besides the insulin cDNA insert) are need to determine which bacterial cells have been transformed and to determine which transformed bacterial cells have a plasmid with the cDNA insulin insert? What is needed in the plasmid with the cDNA insert besides the gene for insulin to actually cause the bacteria to express the insulin gene and produce the insulin protein?arrow_forwardA.) Transformation is best described as: Group of answer choices The integration of foreign DNA into a genome The uptake of naked DNA from the environment The transfer of DNA via a bacteriophage Transfer of a plasmid from one organism to another B.) What is the function of the araC gene in the pGLO plasmid? Group of answer choices It encodes the protein that glows under ultraviolet light It allows us to select for the cells that contain the plasmid It prevents the transcription of the green fluorescent protein unless arabinose is added It ensures that the plasmid will be copied and passed on to daughter cells C.) What is the function of the bla gene in the pGLO plasmid? Group of answer choices It encodes the protein that glows under ultraviolet light It allows us to select for the cells that contain the plasmid It prevents the transcription of the green fluorescent protein unless arabinose is added It ensures that the plasmid will be copied and passed on…arrow_forwardA molecular geneticist hopes to find a gene in human liver cells that codes for an important blood-clotting protein. He knows that the nucleotide sequence of a small part of the gene is CTCGACTCACA. Briefly explain how to obtain the desired gene. Briefly describe how to clone the desired gene into a cloning plasmid.arrow_forward
- What specifically will happen if DNA polymerase is inaccurate during DNA synthesis? Explain how this inaccuracy might affect the organism.arrow_forwarda) Primers can sometimes bind and target the wrong gene, especially if the primers are allowed to bind to the DNA strands at a low temperature. PCR also preferentially amplify short segments of DNA. Would it be important to actually run the cDNA after the PCR on a DNA gel in order to check for a PCR product of the predicted size for the insulin gene? Why or why not?arrow_forwardYou are performing a PCR reaction but unbeknownst to you, there is a significant pool of dUTP in the nucleotide mix (along with dCTP, dTTP, dATP, and dGTP). How might this affect your PCR product? a. If the PCR product was ligated into a plasmid and put into a cell, a totally different mRNA would be made from the insert compared to an insert made with T's. b. If the pool of dTTP ran out before the pool of dUTP, DNA replication could no longer occur. c. During the reaction, uracils incorporated into the product would cause the PCR product to degrade as it is being made. d. Uracil would be incorporated into the product and would lessen the affinity of any DNA binding proteins that might bind to the product in subsequent experiments. e. Nothing would happen since polymerases can't use dUTP to make DNA.arrow_forward
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