Campbell Biology: Custom Edition
18th Edition
ISBN: 9781323717271
Author: Urry, Cain, Wasserman, Minorsky, Reece
Publisher: PEARSON C
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Chapter 20, Problem 11TYU
DRAW IT You are cloning an aardvark gene, using a bacterial plasmid as a vector. The green diagram shows the plasmid, which contains the restriction site for the enzyme used in Figure 20.5. Above the plasmid is a segment of linear aardvark DNA that was synthesized using PCR. Diagram your cloning procedure, and show what would happen to these two molecules during each step. Use one color for the aardvark DNA and its bases and another color for those of the plasmid. Label each step and all 5' and 3' ends.
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Students have asked these similar questions
In biotechnology, gene cloning is a very important technique. A vector is normally
required to perform this process. The vector commonly used to transform a bacterial host
cell is the plasmid. State the three (3) important regions of the plasmid. Elaborate your
answer.
You are studying a new plasmid, and you digest the plasmid with three restriction
enzymes: Eco RI (E), HindlII (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.
E
H
E+H E+X H+X
Kb
+4.3
+2.8
+2.5
-
+2.0
--
-1.8
-1.5
12
+1.0
+0.8
+0.5
-
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?
After transformation you were asked to grow bacterial cells transformed with plasmid on a plate that had X-gal and ampicillin. X-Gal is often used as in indicator dye, which turns blue when metabolized by B-galactosidase protein and used to test if cloning experiments have worked. [Note look at the vector diagrams carefully]
Briefly explain how you would find the bacterial cells that are transformed with the plasmid with the YFG inserted.
Chapter 20 Solutions
Campbell Biology: Custom Edition
Ch. 20.1 - Prob. 1CCCh. 20.1 - DRAW IT One Strand of a DNA molecule has the...Ch. 20.1 - What are some potential difficulties in using...Ch. 20.1 - VISUAL SKILLS Compare Figure 20.7 with Figure...Ch. 20.2 - Prob. 1CCCh. 20.2 - Prob. 2CCCh. 20.3 - Based on current knowledge, how would you explain...Ch. 20.3 - Prob. 2CCCh. 20.3 - Prob. 3CCCh. 20.4 - What is the advantage of using stem cells for gene...
Ch. 20.4 - Prob. 2CCCh. 20.4 - Prob. 3CCCh. 20 - Describe how the process of gene doning results in...Ch. 20 - What useful Information is obtained by detecting...Ch. 20 - Describe how, using mice. a researcher could carry...Ch. 20 - What factors affecf whether a given genetic...Ch. 20 - In DNA technology, the term vector can refer to...Ch. 20 - Which of the following tools of DNA technology is...Ch. 20 - Prob. 3TYUCh. 20 - A paleontologist has recovered a bit of tissue...Ch. 20 - DNA technology has many medical applications....Ch. 20 - Which of the following is not true of cDNA...Ch. 20 - Expression of a cloned eukaryotic gene in a...Ch. 20 - Which Ii of the following sequences in...Ch. 20 - Prob. 9TYUCh. 20 - MAKE CONNECTIONS Looking at Figure 20.15, what...Ch. 20 - DRAW IT You are cloning an aardvark gene, using a...Ch. 20 - EVOLUTlON CONNECTION Ethical considerations aside,...Ch. 20 - Prob. 13TYUCh. 20 - Prob. 14TYUCh. 20 - The water in the Yellowstone National Park hot...
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- Decide on two restriction sites that you can use to clone this into pL4440’s MCS. Identify their sequence. Tip: The plasmid map is in Figure 3, details of restriction site sequences can be found at https://enzymefinder.neb.com/#!#nebheaderarrow_forwardBelow is shown a plasmid map from a recombinant DNA experiment you have performed. You inserted your gene of interest (labeled gene on the map) into the lac Z reporter gene via a Hind III site. If you digested this recombinant DNA with HindIII what is the size of the DNA fragment(s) you would obtain on a DNA gel electrophoresis? The plasmid alone is 4000 bp. I have attached the image below.arrow_forwardYou are asked to sequence a piece of DNA to determine if it is from a gene thought to be involved in the development of breast cancer. The sequence of the template strand is ATGCCCGTAATCGTTA and you are given the primer TAACGA. You take these along with a sequencing kit that contains everything else needed for sequencing. You then run the sequencing experiment and analyze the results on a sequencing gel. Which of the following gels (A-D) is the correct sequencing gel for this experiment? Answers: A-D A A BB CC DD Question #3 attachment A ATOC B с A TO C ATOC ATOCarrow_forward
- The image below shows the general structure of a gene on a chromosome. The arrows above and below the chromosome indicate the binding positions of potential forward (F) and reverse (R) PCR primers. Select two primers from the list below that would exclusively amplify exon 3 in a PCR reaction. ut of Intron 1 Intron 2 +1 Poly-A signal ATG TAG F1 Exon 1 F4 Exon 2 F6 Exon 3 R6 R4 R3 R2 R1 Promoter Select all that apply: cross out Da. F1 cross out Ob. F2 cross out Oc. F3 cross out O d. F4 cross out O e. F5 cross out f. F6 cross out g. R1 cross out Oh. R2 cross out OL R3 cross out O. R4 cross out k. R5 cross out R6 TIarrow_forwardDescribe 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.arrow_forwardYou have set up a recombinant DNA experiment using the plasmid PBR322 as the vector (see plasmid below). You use the BamHI restriction site on the plasmid to insert the target DNA. The plasmid is then used to transform E.coli colls Is the following statement True or False? Growth of the transformed cells on agar containing both ampicillin and tetracycline will eliminate any cells that do not contain a plasmid. Clal Hindlll EcoRI Pvul BamHI Pstl amp tet PBR322 -Sall ori rop Pvull True Falsearrow_forward
- Draw the gel electrophoresis results for the following 16 kb plasmid when the plasmid is cut withrestriction enzyme A, with restriction enzyme B and with both A and B restriction enzymes. The Aenzyme cuts at positions 0 and 9.2 kb and the sites are shown as white. (0 is at the top of the circle.) TheB enzyme cuts at positions 3.8, 7 and 13.2 kb and the sites are shown as gray. Show the electricalpolarity of the gel, the bands, and their sizes in your drawing. Make sizes consistent between gel lanes.Explain why the concept of charge density is important for gel electrophoresis.arrow_forwardOn the gel shown below are four DNA samples. Samples A to C are taken from tissues of landslide victims that are being identified, while sample D came from a hair sample brought by a mother looking for the remains of her son. (see img) i. If similar band patterns in a gel are created using the same restriction enzyme, what does that tell you about the DNA sequence of the samples? ii. In sample C, only two fragments were created. How many restriction sites (regions where enzymes cut) are present in sample C?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_forward
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