Prescott's Microbiology
10th Edition
ISBN: 9781259281594
Author: Joanne Willey, Linda Sherwood Adjunt Professor Lecturer, Christopher J. Woolverton Professor
Publisher: McGraw-Hill Education
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Chapter 17.3, Problem 2MI
What would you conclude if you obtained only blue colonies after cloning into a vector that enables blue/white screening of transformants? Why might such a result occur?
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After mutagenesis of wild type Vibrio fisheri, you isolate two different mutant strains (A and B) that, unlike the wild type cells, fail to luminesce when grown to high density in a flask with appropriate medium. Curiously, however, when you inoculate both mutant strains in the same flask, you observe that the mixed (A+B) culture begins to emit light after growing dense.
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Chapter 17 Solutions
Prescott's Microbiology
Ch. 17.1 - Examine the uncut piece of DNA shown in the upper...Ch. 17.1 - Which of the above enzymes yield blunt ends? Which...Ch. 17.1 - Why might two DNA fragments inadvertently be...Ch. 17.1 - Why must introns be removed from eukaryotic DNA...Ch. 17.1 - If a linear piece of DNA is cut with a restriction...Ch. 17.1 - Describe restriction enzymes, sticky ends, and...Ch. 17.1 - What is cDNA? Why is it necessary to generate cDNA...Ch. 17.1 - You want to visualize a digested plasmid that...Ch. 17.1 - What is the purpose of Southern blotting? How is a...Ch. 17.2 - Why, after three cycles, are the vast majority of...
Ch. 17.3 - Which plasmid is a shuttle vector? Why?Ch. 17.3 - What would you conclude if you obtained only blue...Ch. 17.3 - In what ways does the BAC shown here differ from...Ch. 17.3 - Briefly describe the polymerase chain reaction....Ch. 17.3 - Why is PCR used to detect infectious agents that...Ch. 17.3 - How would you use PCR to measure the concentration...Ch. 17.3 - Why is it possible to visualize a PCR product on...Ch. 17.3 - You want to clone a 6,000 bp DNA fragment in E...Ch. 17.5 - Why are long fragments (e.g., 20,000 bp) of...Ch. 17.6 - What special considerations are necessary if one...Ch. 17.6 - Prob. 1RIACh. 17.6 - Explain the selection for antibiotic resistance...Ch. 17.6 - Prob. 3RIACh. 17.6 - Prob. 4RIACh. 17.6 - Prob. 5RIACh. 17.6 - You are studying chemotaxis proteins in a newly...Ch. 17 - Which of the DNA molecules shown are recombinant?Ch. 17 - You are performing a PCR to amplify a gene...Ch. 17 - Prob. 2CHICh. 17 - Suppose you transformed a plasmid vector carrying...Ch. 17 - You are interested in the activity and regulation...Ch. 17 - Prob. 5CHICh. 17 - Zymomonas mobilis is a Gram-negative bacterium...
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- 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_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_forwardThe temperature at which the primers and target DNA hybridize may be changed to influence the stringency of PCR amplification. What effect will changing the hybridization temperature have on the amplification? Let's say you have a certain yeast gene A and want to check whether it has a human equivalent. How might managing the hybridization's rigor benefit you?arrow_forward
- Consider the following experiment. First, large populations of two mutant strains of Escherichia coli are mixed, each requiring a different, single amino acid. After plating them onto a minimal medium, 45 colonies grew. Which of the following may explain this result? A) The colonies may be due to back mutation (reversion). B) The colonies may be due to recombination. C) Either A or B is possible. D) Neither A nor B is possible.arrow_forwardTransposon 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 -arrow_forwardHow can the desired product formed after genetic engineering be produced on a commercial scale?arrow_forward
- As you are performing this protocol, you realize that the ultraviolet (UV) light does not work as expected, because of a fused bulb. You decide to look at plate #4 (NA with AMP; pGFP added to cells) under visible light to decide whether transformation was successful or not. Will viewing this plate under visible light tell you whether you were successful in transforming cells with pGFP DNA?arrow_forwardWhat would be an advantage of using HaeIII for a cloning experiment? What would be a disadvantage?arrow_forwardFOR PKU: Draw what you’d expect to see if you were to analyze the gene products by conventional PCR, RT-PCR and Western blot. Would you see abnormally sized gene products? Explain and be sure to include any relevant control(s) that you should include in your experiment.arrow_forward
- A person with a rare genetic disease has a sample of her chromosomessubjected to in situ hybridization using a probe that is known to recognize band p11 on chromosome 7. Even though her chromosomes look cytologically normal, the probe does not bind to this person’s chromosomes. How would you explain these results? How would you use this information to positionally clone the gene that is related to this disease?arrow_forwardA technique called fluorescence in situ hybridization (FISH) is described. In this method, a labeled piece of DNA is hybridized to a set of chromosomes. Let’s suppose that you cloned a piece of DNA from G. pubescens and used it as a labeled probe for in situ hybridization. What would you expect to happen if this DNA probe were hybridized to the G. speciosa or G. tetrahit chromosomes? Describe the expected results.arrow_forwardDescribe the outcome of a chain-terminator sequencing procedure in which (a) too little ddNTP is added or (b) too much ddNTP is added.arrow_forward
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