Genetic Analysis: An Integrated Approach (2nd Edition)
2nd Edition
ISBN: 9780321948908
Author: Mark F. Sanders, John L. Bowman
Publisher: PEARSON
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Chapter 17, Problem 3P
Ligase catalyzes a reaction between the
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#3) Ligase catalyzes a reaction between the 5' phosphate and the 3' hydroxyl groups at the end of DNA molecules. The enzyme calf intestinal phosphatase catalyzes the removal of the 5' phosphate from DNA molecules. What would be the consequence of treating a cloning vector, before ligation, with calf intestinal phosphatase?
in the cloning vector, what would be the plausible impacts if a mutation at the Ori site renders it non- functional?
Your cloning vector has restriction recognition sites for two restriction endonucleases, EcorI and BamHI. However, the DNA to be manipulated does not have recognition sites for these two restriction endonucleases. How would you construct a recombinant DNA for the given DNA?
Chapter 17 Solutions
Genetic Analysis: An Integrated Approach (2nd Edition)
Ch. 17 - 15.1 What purpose do the bla and lacZ genes serve...Ch. 17 - The human genome is 3109 bp in length. How many...Ch. 17 - 15.3 Ligase catalyzes a reaction between the...Ch. 17 - You have constructed four different libraries: a...Ch. 17 - Using the genomic libraries in Problem 4, you wish...Ch. 17 - The human genome is 3109bp. You wish to design a...Ch. 17 - 15.7 Using animal models of human diseases can...Ch. 17 - 15.8 Compare methods for constructing homologous...Ch. 17 - 15.9 Chimeric genefusion products can be used for...Ch. 17 - 15.10 Why are diseases of the blood simpler...
Ch. 17 - Injection of double-stranded RNA can lead to gene...Ch. 17 - Compare and contrast methods for making transgenic...Ch. 17 - 15.13 It is often desirable to insert cDNAs into a...Ch. 17 - 15.14 A major advance in the s was the development...Ch. 17 - 15.15 The bacteriophage lambda genome can exist in...Ch. 17 - 15.16 The restriction enzymes Xho and Sal cut...Ch. 17 - 15.17 The bacteriophage has a single-stranded DNA...Ch. 17 - 15.20 You have identified a cDNA clone that...Ch. 17 - You have isolated a genomic clone with an EcoR I...Ch. 17 - 15.18 To further analyze the CRABS CLAW gene (see...Ch. 17 - 15.21 You have isolated another cDNA clone of the...Ch. 17 - 15.22 You have identified five genes in S....Ch. 17 - You have generated three transgenic lines of maize...Ch. 17 - 15.24 Bacterial Pseudomonas species often possess...Ch. 17 - 15.25 Two complaints about some transgenic plants...Ch. 17 - 15.26 In Drosophila, lossoffunction Ultrabithorax...Ch. 17 - Prob. 27PCh. 17 - The highlighted sequence shown below is the one...Ch. 17 - The RAS gene encodes a signaling protein that...Ch. 17 - Vitamin E is the name for a set of chemically...Ch. 17 - 15.31 You have cloned a gene for an enzyme that...
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- How is telomerase related to aging? How can telomerase-related aging beaddressed therapeutically? What is a potential danger of telomerase activation?arrow_forwardWhat is a cloning vector? Give two examples of specific DNA molecules routinely used as cloning vectors.arrow_forwardYou are engineering a new vector that contains a screenable marker that can be used for blue/white screening of successful clones. For each site (1, 2, and 3) on the cloning vector below, describe why it would or would not be a good place for you to put the polylinker to facilitate blue/white screening. You can assume that the polylinker itself will not interfere with coding sequence in that region. In other words, the polylinker length will be a multiple of 3 nucleotides, will not contain a stop codon, and any amino acids translated will not affect the activity of the protein in that region. The arrows indicate the direction of transcription for the gene.arrow_forward
- You are engineering a new vector that contains a screenable marker that can be used for blue/white screening of successful clones. For each site (1, 2, and 3) on the cloning vector below, describe why it would or would not be a good place for you to put the polylinker to facilitate blue/white screening. You can assume that the polylinker itself will not interfere with coding sequence in that region. In other words, the polylinker length will be a multiple of 3 nucleotides, will not contain a stop codon, and any amino acids translated will not affect the activity of the protein in that region. The arrows indicate the direction of transcription for the gene. Note from student:As stated in the problem... "YOU CAN ASSUME THAT THE POLYLINKER ITSELF WILL NOT INTERFERE WITH CODING SEQUENCE IN THAT REGION. IN OTHER WORDS, THE POLYLINKER LENGTH WILL BE A MULTIPLE OF 3 NUCLEOTIDES, WILL NOT CONTAIN A STOP CODON, AND ANY AMINO ACIDS TRANSLATED WILL NOT AFFECT THE ACTIVITY OF THE PROTEIN IN THAT…arrow_forwardWhich restriction enzyme is best suited for cloning in pUC8?arrow_forwardWhat is linker scanning mutagenesis?arrow_forward
- The plasmid cloning vector pBR322 is cleaved with the restriction endonuclease PstI. An isolated DNA fragment from a eukaryotic genome (also produced by PstI cleavage) is added to the prepared vector and ligated. The mixture of ligated DNAs is then used to transform bacteria, and plasmid-containing bacteria are selected by growth in the presence of tetracycline. The cloned DNA fragment is 1,000 bp long and has an EcoRI site 250 bp from one end. Three different recombinant plasmids are cleaved with EcoRI and analyzed by gel electrophoresis, giving the patterns shown below. What does each pattern say about the cloned DNA? Note: pBR322, the PstI and EcoRI restriction sites are about 750 bp apart. The entire plasmid with no cloned insert is 4,361 bp. Size markers in lane 4 have the number of nucleotides noted.arrow_forwardEarly gene-cloning experiments involved insertion at one restriction site in the vector; for example, the insert would have an EcoRI site at each end, and the vector would be opened at an EcoRI site prior to ligation. Under what circumstances would asymmetric cloning be desirable, with the insert having a different restriction site at each endarrow_forwardWhat is an STS? How are STSs generated experimentally? What are the uses of STSs? Explain how a microsatellite can be a polymorphic STS.arrow_forward
- In producing genetically engineered human insulin in bacteria, why is it important to use the samerestriction enzyme to cut both the human DNA and the bacterial plasmid?arrow_forwardA 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 bparrow_forwardKnowing that you are using HindIII and EcoRI to cut your plasmids, and that those two enzymes cut within the MCS, use the map of pUC19 provided below to compute: What will be the sizes of the 2 restriction fragments if NO insert is present in pUC19? What will be the sizes of the 2 restriction fragments if the approximately 317 bp RT-PCR product (insert from WT satC dimer) was ligated successfully into the SmaI site? What will be the sizes of the 2 restriction fragments if TWO approximately 317 bp RT-PCR products (2 ligated inserts from WT satC dimer) were ligated into the SmaI site?arrow_forward
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