Genetic Analysis: An Integrated Approach (3rd Edition)
3rd Edition
ISBN: 9780134605173
Author: Mark F. Sanders, John L. Bowman
Publisher: PEARSON
expand_more
expand_more
format_list_bulleted
Concept explainers
Videos
Textbook Question
Chapter 15, Problem 31P
You have cloned a gene for an enzyme that degrades lipids in a bacterium that normally lives in cold temperatures. You wish to transfer this gene into E. coli to produce industrial amounts of enzyme for use in laundry detergent.
How would you accomplish this?
You have managed to produce transgenic E. coli expressing mRNA of your gene, but only a low level of protein is produced. Why might this be so? How could you overcome this problem?
Expert Solution & Answer
Want to see the full answer?
Check out a sample textbook solution
Students have asked these similar questions
Bacteria can be used to produce human growth hormone (HGH - a peptide/protein) through genetic engineering. The human gene for HGH is inserted into a plasmid, which is then
taken up by a bacterial cell, which divides and multiplies into a clone of cells, all of which contain the plasmid with the HGH gene. The bacteria express the HGH gene, producing
HGH which can be harvested and used for treatment of humans. (See figure below) Which of the following statements is NOT true about this process?
bacterium
Vector, such as a
DNA containing the gene of
plasmid, isolated
it from a different species is
Gene encoding protein for
pest resistance is inserted
into plant cells
©2019 Pearson Education, Inc
chromosome recombinant
DNA (plasmid)
transformed
bacterium
Create and harvest
copies of a gene
with either of two goals in mind.
Gene encoding degradative
enzyme to clean up toxdo
waste is inserted into
bacterial cells
ved by an enzyme into
gene of interest
The desired gene is selected and…
Zoey Wong is a research officer at the Department of Biosciences of Tunku Abdul Rahman
University College. Her supervisor instructs her to prepare chemically competent cells for
heat shock transformation from old batches of competent cells that are already available in the
freezers. The competent cells will eventually be used for the expression of a prokaryotic
enzyme using the pET vector system.
(i)
(ii)
You found several different strains of Escherichia coli in the -80 C freezer. State
three (3) E. coli strains you would use for your project which involves the cloning
and expression of the protein of interest.
In order for cloning and expression to happen optimally, suggest two (2) types of pET
vectors suitable for your project.
Transposon mutagenesis was used to generate a library of
mutants within the Salmonella genome. You are trying to
identify a colony with the transposon inserted in the
pathogenic related gene SPI-1 using PCR. Forward and
reverse primers are generated that flank either side of the
gene and yield a wild type product that is 900 bases in
length. Which of the colonies sampled in the gel would
you expect to contain the SPI-1 gene with transposon
insertion?
3,000
2,000
1,000
700
500
300
100
Ladder Colony A Colony B Colony C Colony D Colony E
none
colonies A&C
colonies B&E
O colonies A, C, &D
colonies B, D, &E
-
Chapter 15 Solutions
Genetic Analysis: An Integrated Approach (3rd Edition)
Ch. 15 - 15.1 What purpose do the bla and lacZ genes serve...Ch. 15 - The human genome is 3109 bp in length. How many...Ch. 15 - 15.3 Ligase catalyzes a reaction between the...Ch. 15 - You have constructed four different libraries: a...Ch. 15 - Using the genomic libraries in Problem 4, you wish...Ch. 15 - The human genome is 3109bp. You wish to design a...Ch. 15 - 15.7 Using animal models of human diseases can...Ch. 15 - 15.8 Compare methods for constructing homologous...Ch. 15 - 15.9 Chimeric genefusion products can be used for...Ch. 15 - 15.10 Why are diseases of the blood simpler...
Ch. 15 - Injection of double-stranded RNA can lead to gene...Ch. 15 - Compare and contrast methods for making transgenic...Ch. 15 - 15.13 It is often desirable to insert cDNAs into a...Ch. 15 - 15.14 A major advance in the s was the development...Ch. 15 - 15.15 The bacteriophage lambda genome can exist in...Ch. 15 - 15.16 The restriction enzymes Xho and Sal cut...Ch. 15 - 15.17 The bacteriophage has a single-stranded DNA...Ch. 15 - 15.18 To further analyze the CRABS CLAW gene (see...Ch. 15 - You have isolated a genomic clone with an EcoR I...Ch. 15 - 15.20 You have identified a cDNA clone that...Ch. 15 - 15.21 You have isolated another cDNA clone of the...Ch. 15 - 15.22 You have identified five genes in S....Ch. 15 - You have generated three transgenic lines of maize...Ch. 15 - 15.24 Bacterial Pseudomonas species often possess...Ch. 15 - 15.25 Two complaints about some transgenic plants...Ch. 15 - 15.26 In Drosophila, lossoffunction Ultrabithorax...Ch. 15 - Prob. 27PCh. 15 - The highlighted sequence shown below is the one...Ch. 15 - Vitamin E is the name for a set of chemically...Ch. 15 - The RAS gene encodes a signaling protein that...Ch. 15 - 15.31 You have cloned a gene for an enzyme that...Ch. 15 - 15.32 About of occurrences of nonautoimmune type...Ch. 15 - Describe how having the Cas 9 gene at a genomic...Ch. 15 - 15.34 Would a gene drive system spread rapidly...
Knowledge Booster
Learn more about
Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.Similar questions
- You want to clone a eukaryotic gene and express the corresponding protein in yeast. However, the protein typically localizes within mitochondria. How will you perform your gene cloning so that the protein is secreted from the cell, rather than localized within yeast mitochondria?arrow_forwardThe human hexokinase enzyme has the same function as the bacterial hexokinase enzyme but is somewhat different in its amino acid sequence. You have obtained a mutant bacterial strain in which the gene for hexokinase is missing. If you introduce into your mutant strain a DNA plasmid engineered to contain the DNA coding sequence of the human hexokinase gene, what must you also include? a)The human hexokinase promoter b)The bacterial hexokinase promoter c)Both the human and bacterial promoters d)You cannot engineer a bacteria to produce a human enzymearrow_forwardGenetic transfer via transformation can also be used to map genes along the bacterial chromosome. In this approach, fragments of chromosomal DNA are isolated from one bacterial strain and used to transform another strain. The experimenter examines the transformed bacteria to see if they have incorporated two or more different genes. For example, the DNA may be isolated from a donor E. coli bacterium that has functional copies of the araB and leuD genes. Let’s call these genes araB+ and leuD+ to indicate the genes are functional. These two genes are required for arabinose metabolismand leucine synthesis, respectively. To map the distance betweenthese two genes via transformation, a recipient bacterium is used that is araB− and leuD−. Following transformation, the recipient bacterium may become araB+ and leuD+. This phenomenon is calledcotransformation because two genes from the donor bacterium have been transferred to the recipient via transformation. In this type of experiment, the…arrow_forward
- Genetic transfer via transformation can also be used to map genes along the bacterial chromosome. In this approach, fragments of chromosomal DNA are isolated from one bacterial strain and used to transform another strain. The experimenter examines the transformed bacteria to see if they have incorporated two or more different genes. For example, the DNA may be isolated from a donor E. coli bacterium that has functional copies of the araB and leuD genes. Let’s call these genes araB+ and leuD+ to indicate the genes are functional. These two genes are required for arabinose metabolismand leucine synthesis, respectively. To map the distance betweenthese two genes via transformation, a recipient bacterium is used that is araB− and leuD−. Following transformation, the recipient bacterium may become araB+ and leuD+. This phenomenon is calledcotransformation because two genes from the donor bacterium have been transferred to the recipient via transformation. In this type of experiment, the…arrow_forwardGenetic transfer via transformation can also be used to map genes along the bacterial chromosome. In this approach, fragments of chromosomal DNA are isolated from one bacterial strain and used to transform another strain. The experimenter examines the transformed bacteria to see if they have incorporated two or more different genes. For example, the DNA may be isolated from a donor E. coli bacterium that has functional copies of the araB and leuD genes. Let’s call these genes araB+ and leuD+ to indicate the genes are functional. These two genes are required for arabinose metabolismand leucine synthesis, respectively. To map the distance betweenthese two genes via transformation, a recipient bacterium is used that is araB− and leuD−. Following transformation, the recipient bacterium may become araB+ and leuD+. This phenomenon is calledcotransformation because two genes from the donor bacterium have been transferred to the recipient via transformation. In this type of experiment, the…arrow_forwardThe following DNA sequence is from a bacteriophage that infects a pathogenic bacterium and scientists want to know if this bacteriophage could prove to be a potential treatment against it. But first scientists need to discover if different strains of this pathogen have restriction endonucleases that it may use for its own protection. They try 3 different RE’s:a) EcoR1 b) HaeIII c) BamH1 Look up the recognition sequences for the 3 Res. Enzymes above and check whether the phage genome (a snippet of which is shown below) will or will not be ‘cut’. Tell me how their experiment worked out and what their conclusion was.G A A A A G G C C A C A A G G C C G T C G A C T T T T A A A A G G C C A C A T G C G G C T T T T C C G G T G T T C C G G C AG C T GA A A AT T T T C C G G T G T A C G CCarrow_forward
- You’ve made your construct and placed it into E. coli! Congratulations, you have made a transgenic organism. Your investors will want to know about quality control. How will you check that the correct piece of DNA is in your vector? How will you check to make sure the gene is transcribed? How will you check to make sure that the GasP protein is made in E. coli? Your investors are concerned that the GasP protein might not be sufficiently produced under normal laboratory conditions. They suggest controlling the transcription of the gasP gene using a chemical that will “trigger” its transcription. What type of promoter could be used? What chemical will you use to control transcription? How does this method of control work?arrow_forwardWe have two specific strains of E. coli that have shown horizontal gene transfer (HGT) when mixed. To experimentally determine the method of HGT that is happening, the following conditions are set up in different tubes of culture media: A) Donor and recipient strain mixed together (control - no treatment). B) Donor and recipient strains mixed together, DNase added (can digest DNA in solution, not within cells).C) Special tube containing a membrane filter (with pores that allow DNA and viruses to pass through, but not bacterial cells) that separates two compartments. Donor strain is added on one side, the recipient strain on the other (they are separated by the filter).D) Donor and recipient strains mixed together, with chemical that inactivates viruses (chemical affects bacteriophages in solution so they are unable to attach to cells). The results: Tubes A, B, and D: HGT was observed. Tube C: HGT was NOT observed. Based on this, which type of HGT was occurring? Conjugation,…arrow_forwardArrange the steps that would be used in a laboratory to engineer a bacterium that could express the human gene coding for factor VIII. Not all steps will be placed. Isolate the human gene that produces factor VIII. Isolate the mRNA of the factor VIII gene. Generate cDNA of the factor VIII gene using reverse transcriptase. Incorrect Insert the factor VIII cDNA into a bacterial vector near a promoter site. Transform the vector into an E. coli bacterium. Human DNA is introduced into the bacterial cell using direct injection by a small needle. E. coli expresses factor VIII. Answer Bank The factor VIII protein is isolated from human tissues.arrow_forward
- A research group is studying a bacterium X that binds to mucosal cells in the lung and invades. Wildtype X has an LD50 value of 10 bacteria when administered to mice by inhalation. Using transposon mutagenesis, the researchers have isolated two mutants of X that they call Xmut1 and Xmut2, both of which have LD50 values of 105 when inhaled by mice. However, in tissue culture cells, Xmut1 can invade the cells just as well as wild-type X, while Xmut2 cannot. Provide a possible explanation for these results.arrow_forwardYou want to clone a eukaryotic gene and express the protein in yeast. The protein typically localizes in the mitochondria. What does your clone need so that the protein is localized in the ER rather than the mitochondria?arrow_forwardMutations in the CFTR gene result in cystic fibrosis in humans, a conditions in which abnormal secretions are present in the lungs, pancreas, and sweat glands. The gene was mapped to a 500-kb region on chromosome 7 containing 3 candidate genes. a)Using your knowledge of the disease symptoms, how would you distinguish between the candidate genes to decide which is most likely to encode the CFTR gene? b)How would you prove that your chosen candidate is the CFTR gene?arrow_forward
arrow_back_ios
SEE MORE QUESTIONS
arrow_forward_ios
Recommended textbooks for you
Human Anatomy & Physiology (11th Edition)BiologyISBN:9780134580999Author:Elaine N. Marieb, Katja N. HoehnPublisher:PEARSON
Biology 2eBiologyISBN:9781947172517Author:Matthew Douglas, Jung Choi, Mary Ann ClarkPublisher:OpenStax
Anatomy & PhysiologyBiologyISBN:9781259398629Author:McKinley, Michael P., O'loughlin, Valerie Dean, Bidle, Theresa StouterPublisher:Mcgraw Hill Education,
Molecular Biology of the Cell (Sixth Edition)BiologyISBN:9780815344322Author:Bruce Alberts, Alexander D. Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter WalterPublisher:W. W. Norton & Company
Laboratory Manual For Human Anatomy & PhysiologyBiologyISBN:9781260159363Author:Martin, Terry R., Prentice-craver, CynthiaPublisher:McGraw-Hill Publishing Co.
Inquiry Into Life (16th Edition)BiologyISBN:9781260231700Author:Sylvia S. Mader, Michael WindelspechtPublisher:McGraw Hill Education
Human Anatomy & Physiology (11th Edition)
Biology
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:PEARSON
Biology 2e
Biology
ISBN:9781947172517
Author:Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:OpenStax
Anatomy & Physiology
Biology
ISBN:9781259398629
Author:McKinley, Michael P., O'loughlin, Valerie Dean, Bidle, Theresa Stouter
Publisher:Mcgraw Hill Education,
Molecular Biology of the Cell (Sixth Edition)
Biology
ISBN:9780815344322
Author:Bruce Alberts, Alexander D. Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter Walter
Publisher:W. W. Norton & Company
Laboratory Manual For Human Anatomy & Physiology
Biology
ISBN:9781260159363
Author:Martin, Terry R., Prentice-craver, Cynthia
Publisher:McGraw-Hill Publishing Co.
Inquiry Into Life (16th Edition)
Biology
ISBN:9781260231700
Author:Sylvia S. Mader, Michael Windelspecht
Publisher:McGraw Hill Education
Molecular Techniques: Basic Concepts; Author: Dr. A's Clinical Lab Videos;https://www.youtube.com/watch?v=7HFHZy8h6z0;License: Standard Youtube License