Microbiology: An Introduction (13th Edition)
13th Edition
ISBN: 9780134605180
Author: Gerard J. Tortora, Berdell R. Funke, Christine L. Case, Derek Weber, Warner Bair
Publisher: PEARSON
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Chapter 9, Problem 2CAE
Using the restriction enzyme ECORI, the following gel electrophoresis patterns were obtained from digests of various DNA molecules from a transformation experiment. Can you conclude from these data that transformation occurred? Explain why or why not.
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A 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?
A 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.
You are trying to clone a gene. You have successfully isolated it from the genomic DNA of an organism using
the Hindlll restriction enzyme. You then take a plasmid with a single EcoRI restriction site and cleave it with
EcoRI. You combine these two fragments and treat them with DNA ligase. Answer the two questions below.
a.(2 points Does the cloning reaction succeed as described? If so, what is the product obtained?
b.
Explain your answer above.
Chapter 9 Solutions
Microbiology: An Introduction (13th Edition)
Ch. 9 - Compare and contrast the following terms: a. cDNA...Ch. 9 - Differentiate the following terms. Which one is...Ch. 9 - Some commonly used restriction enzymes are listed...Ch. 9 - Suppose you want multiple copies of a gene you...Ch. 9 - Which enzyme makes the smallest fragment...Ch. 9 - Describe a recombinant DNA experiment in two or...Ch. 9 - List at least two examples of the use of rDNA in...Ch. 9 - You are attempting to insert a gene for saltwater...Ch. 9 - How does RNAi silence a gene?Ch. 9 - Prob. 10R
Ch. 9 - Restriction enzymes were first discovered with the...Ch. 9 - The DNA probe, 3-GGCTTA, will hybridize with which...Ch. 9 - Which of the following is the fourth basic step to...Ch. 9 - The following enzymes are used to make cDNA. What...Ch. 9 - If you put a gene in a virus, the next step in...Ch. 9 - You have a small gene that you want replicated by...Ch. 9 - Pieces of human DNA stored in yeast cells. a....Ch. 9 - A population of cells carrying a desired plasmid....Ch. 9 - Self-replicating DNA for transmitting a gene from...Ch. 9 - A gene that hybridizes with mRNA. a. antisense b....Ch. 9 - Design an experiment using vaccinia virus to make...Ch. 9 - Why did the use of DNA polymerase from the...Ch. 9 - The following picture shows bacterial colonies...Ch. 9 - Prob. 1CAECh. 9 - Using the restriction enzyme ECORI, the following...
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- 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.arrow_forwardDNA can be cut at specific sequence motifs using enzymes called restriction endonucleases. You obtain a single PCR product from R. sphaeroides DNA that can be cut in the places shown below by solid arrows. These are the Clal specific restriction motifs. Positions of these restriction sites are indicated by nucleotide numbers in a 1023 nucleotide DNA fragment. Draw the expected band(s) for each of the bacterial species on the gel picture below if treated with the restriction enzymes as illustrated. Rhodobacter sphaeroides 2. 4. 1 0. 1023 150 627 Rhodopseudomonas palustris BisB5 0. 1023 627 Dinoroseobacter shibae DFL 12 1023 150 Silicibacter sp. TM 1040 1023 DNA ladder R sphaeroides 2. 4. 1 R palustris BisB5 D. shibae DFL12 Silicibacter sp. TM1040 1100 1000 900 800 700 600 500 400 300 200 100arrow_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.arrow_forward
- Suppose that a geneticist discovers a new restriction enzyme in the bacterium Aeromonas ranidae. This restriction enzyme is the first to be isolated from this bacterial species. Using the standard convention for abbreviating restriction enzymes, give this new restriction enzyme a name (arrow_forwardSuppose that a geneticist discovers a new restriction enzyme in the bacterium Aeromonas ranidae. This restriction enzyme is the first to be isolated from this bacterial species. Using the standard convention for abbreviating restriction enzymes, give this new restriction enzyme a name (for help, see footnote to Table 19.1).arrow_forwardAssume that a plasmid is 4700 base pairs in length and has restriction sites for a given restriction enzyme at the following locations: 800, 1400, 2900, and 3600. List the fragments by size that are ! expected when the plasmid is fully digested the restriction enzyme.arrow_forward
- Construct a detail linear restriction map according to the given information in Table 1 below. Table 1 Restriction enzyme EcoRI No, of fragments 2 Size of fragment (kb) 2,9 ВатHI 3 6, 1,4 Xhol 8, 3 EcoRI + BamHI 4 2, 4, 1,4 EcoRI + Xhol 3 2, 6, 3arrow_forwardRestriction endonuclease digestion of a DNA sequence yielded fragments of the following sizes: 1. 5.2 kb 2. 0.8 kb 3. 1.2 kb 4. 3.8 kb 5. 3.1 kb After gel electrophoresis, what would be the order in which these fragments would be found—the last fragment listed being furthest from the negative pole.arrow_forwardBacterial systems serve as an excellent model to express proteins but has a disadvantage – what is that disadvantage? List the points to differentiate three classes of restriction enzymes? Give an example of restriction enzyme that has ability to generate blunt and cohesive ends after digestion of DNA?arrow_forward
- See the restriction enzyme map below. The total DNA length is 1800 base pairs. If this DNA is cut using three restriction enzymes, namely Kpnl, Sall and EcoRI, it yields four fragments with sizes of 390 bR. 810 bp, 270 bp www and 330 bp. Kpnl Sall EcoRI 390 810 270 330 1800 bp 1. If you were to subject this digested DNA to agarose gel electrophoresis, what would your gel look like? Draw a detailed picture of your gel. Remember to indicate the direction in which your DNA is moving and also show any reference samples. Also remember to show all components of your gel. 2. You are provided with coiled DNA and plasmid DNA that you subject to gel electrophoresis. Draw this gel. Remember to indicate the direction in which your DNA is moving and also show any reference samples. Also remember to show all components of your gel. Exac fragment sizes are not important.arrow_forwardSingle and double digestion of plasmid pMCS326 were performed using the restriction enzymes AluIII and EcoRV. DNA fragments were shown in an electrophoretogram below. Construct a restriction map of plasmid pMCS326 for enzymes AluIII and EcoRV. (Create restriction mapping with explanation)arrow_forwardIn making recombinant DNA molecules that combine restriction fragments from different organisms, researchers usually prefer restriction enzymes like BamHI or HindIII that generate fragments with “sticky ends” (ends with overhangs) rather than enzymes like HpaI or SmaI (Table 12.1) that generate fragments with “blunt ends” (ends without overhangs). Can you think of a reason for this preference?arrow_forward
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