Campbell Biology, Books a la Carte Plus Mastering Biology with eText -- Access Card Package (10th Edition)
10th Edition
ISBN: 9780133922851
Author: Jane B. Reece, Lisa A. Urry, Michael L. Cain, Steven A. Wasserman, Peter V. Minorsky, Robert B. Jackson
Publisher: PEARSON
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Chapter 20.1, Problem 3CC
What are some potential difficulties in using plasmid vectors and bacterial host cells to produce large quantities of proteins from cloned eukaryotic genes?
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In bacterial transformation, the purpose of having antibiotic within an agar plate is to:
Select one:
confirm which plasmids been have successfully ligated with a gene of interest.
isolate bacteria which have been successfully transformed with the plasmid.
indicate which plasmids were successfully digested by the endonuclease.
act as a substrate which will be cleaved and produce a blue product when ligation is unsuccessful.
show which plasmids contain the lacZ gene.
A shuttle vector is a vector (usually a plasmid) constructed so that it can propagate in two
different host species. One of the most common types of shuttle vectors is the
yeast shuttle
vector. Examples of such vectors derived from yeast are Yeast Episomal Plasmid (YEP),
Yeast Integrating Plasmid (YIP) and Yeast Replicating Plasmid (YRP). Why is YEP preferred
over YIP and YRP? Give your thoughts on this.
If you use the pUC18 vector to clone in the MCS region, predict the following:
a) Do bacteria that are blue in color have a cloned insert?
b)Do bacteria that are white in color have a cloned insert?
c) If you were to grow these cells on Chloramphenicol (an antibiotic), would the bacteria with the pUC plasmid grow? Why or Why not?
Chapter 20 Solutions
Campbell Biology, Books a la Carte Plus Mastering Biology with eText -- Access Card Package (10th Edition)
Ch. 20.1 - Prob. 1CCCh. 20.1 - Prob. 2CCCh. 20.1 - What are some potential difficulties in using...Ch. 20.1 - Prob. 4CCCh. 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 - Prob. 5TYUCh. 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 - Prob. 10TYUCh. 20 - EVOLUTlON CONNECTION Ethical considerations aside,...Ch. 20 - Prob. 12TYUCh. 20 - Prob. 13TYUCh. 20 - The water in the Yellowstone National Park hot...
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- A) Outline the experimental procedure for cloning a eukaryotic gene and expressing it in E. coli. Focus on the essential steps starting with eukaryotic gene amplification to transformation of E. coli cells B) Explain how insertional inactivation can help you identify the colonies that carry the plasmid with your eukaryotic gene of interest C) Plasmids containing antibiotic resistance genes are widely used in gene cloning and other molecular biology techniques. What would happen if the eukaryotic gene was inserted into an antibiotic resistance gene on the plasmid?arrow_forwardA shuttle vector is a vector (usually a plasmid) constructed so that it can propagate in two different host species. One of the most common types of shuttle vectors is the yeast shuttle vector. Examples of such vectors derived from yeast are Yeast Episomal Plasmid (YEp), Yeast Integrating Plasmid (YIp) and Yeast Replicating Plasmid (YRp). Among these three vectors, YIp has the lowest transformation frequency and copy number per cell. Explain why Ylp is still popularly used despite its limitations.arrow_forwardWhen using a conventional plasmid cloning vector containing a b-galactosidase gene, it is possible to perform a "blue-white screen" to determine which bacteria have taken up a plasmid into which a DNA fragment as been inserted, as opposed to those that have taken up just reclosed plasmid vector, by growing the transformed cells on nutrient agar plates containing the artificial b-gal substrate X-gal. Will bacteria that have taken up a plasmid into which a DNA fragment has been inserted form a blue colony or a white colony when grown on this medium? Briefly explain why these bacteria would form a colony of the color you chose.arrow_forward
- Plasmids are the only vectors currently available for use in recombinant procedures.True or false?arrow_forwardWhen an E. coli donor cell duplicates a strand of plasmid DNA, and passes this DNA strand to a recipient E. coli cell, without the use of naked DNA in solution or of a viral vector, this is: an example of horizontal gene transfer by means of lysogenic bacteriophages an example of horizontal gene transfer by means of lytic bacteriophages an example of horizontal gene transfer by means of transformation an example of horizontal gene transfer by means of transduction an example of horizontal gene transfer by means of conjugationarrow_forwardList the major steps that would be required to clone the human insulin gene into a plasmid and to identify bacterial colonies that contain the plasmid with the insulin gene inserted. Be sure to include a suggestion for how to select colonies with a plasmid and how to distinguish plasmids with and without the insulin gene insertedarrow_forward
- If we were to examine a strain with the F plasmid inserted into the same site of the bacterial chromosome, but in the reverse orientation: a) What would the order of gene transfer be? Include all of the genetic markers including the amino acid and nucleic acid metabolism genes and streptomycin resistance. b) What cell types would be able to grow on the NA vs ECM media types? Be sure to include the genotypes of the cells that would grow. Remember that NA provides all nutrients the bacteria needs + no antibiotic and HCM = minimal medium + glucose + has streptomycin antibiotic c) Would we still be able to perform our mapping? Why or why not? (Hint: refer to part b above)arrow_forwardHow can I develop a vector to express a bacterial gene (2,500 bp in size) in E. coli. How do I know if the gene of interest is in the vector? What method can I use to get the vector into E. coli?arrow_forwardDescribe the molecular features of a BAC cloning vector. What is theprimary advantage of a BAC vector over a plasmid or viral vector?arrow_forward
- If a vector containing the desired gene X is inserted into a microorganism, for instance, A.carbornarious, and the protoplast transformation resulted in 9 gene X transformants, how to determine which transformant has the highest titer of gene X?arrow_forwardWhen cloning a foreign DNA fragment into a plasmid, it is often useful to insert the fragment at a site that interrupts a selectable marker(such as the tetracycline-resistance gene of pBR322). The loss of function of the interrupted gene can be used to identify clones containing recombinant plasmids with foreign DNA. With a yeast artificial chromosome (YAC) vector, it is not necessary to do this; the researcher can still distinguish vectors that incorporate large foreign DNA fragments from those that do not. How are these recombinant vectors identified?arrow_forwardWith reference to the image below, discuss the process and principle involved for screening/selection of hosts (last stage of cloning) containing the intended recombinant plasmid. LacZ' = Gene for alpha-peptide of β-galactosidase.arrow_forward
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Bacterial Genomics and Metagenomics; Author: Quadram Institute;https://www.youtube.com/watch?v=_6IdVTAFXoU;License: Standard youtube license