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 2IQ
The following schematic diagram depicts an overview of gene cloning. Identify components a-g. Briefly describe what is happening in the three numbered steps of the process. For what two purposes might the bacteria produced in step 3 be used?
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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:
d. 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.
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.
After cloning is carried out to insert a foreign gene into BL21(DE3), you would like to
confirm the expression of the foreign protein in the bacteria using a blotting technique.
Briefly describe the steps to achieve your objective with simple illustrations and descriptions.
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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- The enzymes mentioned below are used as tools during cloning, DNA sequencing and/or gene therapy. Explain what they are used for. Also mention the actual biological function of the respective enzymes. 1) RNaseHarrow_forward#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?arrow_forwardDescribe the various types of vectors used in DNA cloning. Explain the difference between a cloning vector and an expression vector.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.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. 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_forwardDescribe the important features of cloning vectors. Explain the purpose of selectable markers in cloning experiments.arrow_forward
- Cloning vectors are not just limited to bacterial plasmids. Bacteriophages and M13 phage vectors are also commonly utilized in the cloning process. State any five (5) key criteria to be an effective cloning vector.arrow_forwardTranscriptome analysis involves two separate methodologies: gene expression and RNA seq analyses. The 10 items below are a scrambled listing of the steps used in the two procedures. Identify the steps involved in RNA seq from the list below. Use the numbers in the list to refer to each step. Once the steps for RNA seq have been identified, write the steps in the order in which they are performed during the experiment. (1) DNA sequencing (2) Allow for hybridization and wash excess cRNA. (3) Mix labeled cRNA with array chip. (4) PCR amplification (5) Measure fluorescence intensity to determine abundance of transcripts. (6) Add labeled cRNA at each microarray location. (7) Map cDNA sequences to the genome of the organism to determine identity and abundance of transcripts. (8) mRNA isolation from cells (9) Prepare fluorescently labeled cRNA probes (10) cDNA synthesisarrow_forwardWhat does gene shuffling mean, how is it done? What is it used for? What advantages does it have in biotechnology? Explain briefly.arrow_forward
- Relative to this image of the steps in gene cloning, identify the steps involved and the protein/enzyme requirements of each.arrow_forwardWhat is a cloning vector? Give two examples of specific DNA molecules routinely used as cloning vectors.arrow_forwardWhat are the following features that are required facilitate cloning into a vector.arrow_forward
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