Campbell Biology
12th Edition
ISBN: 9780135188743
Author: Urry
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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What are some potential difficulties in using plasmid vectors and bacterial host cells to produce large quantitiesof proteins from cloned eukaryotic genes?
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.
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
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 - 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. 8TYUCh. 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. 12TYUCh. 20 - Prob. 13TYUCh. 20 - Prob. 14TYU
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- 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.arrow_forwardA) 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_forwardWhat characteristics of plasmids makes them good cloning vectors? Discuss.arrow_forward
- 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). 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 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_forward1)Find a plasmid map for pET11a and create a basic procedure for cloning a gene into this vector. Which selection method and substance would you use for this plasmid after transformation? 2) Which organism would be your first choice for transformation: a bacterium, earthworm, fish or mouse? Describe your reasoning. 3) What is the main difference between plates before and after centrifugation? What may be the purpose of concentrating the media and perform a second plating?arrow_forward
- When 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_forwardPlease DESCRIBE, in outline form, the method you will use to select for bacterial cells that have taken up the pL311 plasmid, and to screen those cells for the presence of plasmids that are likely to contain a cloned gene. Be sure to mention the specific media you will use. In addition, please explain the rationale behind this specific selection and screening procedure. (Remember that you have available the following types of media: (i) media containing neither kanamycin nor X-gal, (ii) media containing BOTH kanamycin and X-gal, (iii) media containing tetracycline, and (iv) media containing ampicillin.arrow_forwardPlasmids are the only vectors currently available for use in recombinant procedures.True or false?arrow_forward
- What are the components needed for the processes of transformation, conjugation, and transduction? How does each process occur? What genes are involved in each process? How do generalized and specialized transduction differ? What is the end result of each?arrow_forwardWhen cloning a piece of DNA, the purpose of using the LacZ blue-white colony method is to A) Remove bacteria that do not have recombinant vectors B) Remove bacteria that do not have the DNA insert of interest Remove linear DNA Distinguish colonies that have recombinant vectors from those with non-recombinant vectors. E) Remove bacteria that have not taken up the vectorarrow_forwardIf 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_forward
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