Prescott's Microbiology
11th Edition
ISBN: 9781260211887
Author: WILLEY, Sandman, Wood
Publisher: McGraw Hill
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Chapter 17, Problem 4AL
You are interested in the activity and regulation of a protease made by the Gram-positive bacterium Geobacillus stearothermophilus. What would be the purpose of constructing each of the following: a His-tagged protease, a transcriptional GFP fusion to the protease gene, and a translational GFP fusion to the protease gene?
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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 - Prob. 3MICh. 17.1 - What would you conclude if you obtained only blue...Ch. 17.1 - Why must introns be removed from eukaryotic DNA...Ch. 17.1 - Which plasmid is a shuttle vector? Why?Ch. 17.1 - In what ways does the BAC shown here differ from...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 - Prob. 3CC
Ch. 17.1 - Prob. 4CCCh. 17.1 - Prob. 5CCCh. 17.2 - Why, after three cycles, are the vast majority of...Ch. 17.2 - Briefly describe the polymerase chain reaction....Ch. 17.2 - Why is PCR used to detect infectious agents that...Ch. 17.2 - How would you use PCR to measure the concentration...Ch. 17.2 - Why is it possible to visualize a PCR product on...Ch. 17.2 - Prob. 5CCCh. 17.3 - Why are long fragments (e.g., 20,000 bp) of...Ch. 17.4 - What special considerations are necessary if one...Ch. 17.4 - Prob. 1CCCh. 17.4 - Prob. 2CCCh. 17.4 - Prob. 3CCCh. 17.4 - You are studying chemotaxis proteins in a newly...Ch. 17.5 - Prob. 1MICh. 17.5 - Prob. 1CCCh. 17.5 - Prob. 2CCCh. 17 - Which of the DNA molecules shown are recombinant?Ch. 17 - Prob. 1RCCh. 17 - Prob. 2RCCh. 17 - Prob. 3RCCh. 17 - Prob. 4RCCh. 17 - Prob. 5RCCh. 17 - Prob. 6RCCh. 17 - Prob. 1ALCh. 17 - Prob. 2ALCh. 17 - Suppose you transformed a plasmid vector carrying...Ch. 17 - You are interested in the activity and regulation...Ch. 17 - Prob. 5ALCh. 17 - Prob. 6ALCh. 17 - Prob. 7AL
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- Mabelle used the pET vector system to express her prokaryotic amylase enzyme. She added IPTG into her culture broth of DH5a Escherichia coli strain. At the end of the experiment, she discovered that her protein was not expressed. She repeated three more times but her protein of interest was still not produced. (i) (ii) Explain the reason why Mabelle failed to obtain her protein of interest and suggest a solution to troubleshoot this problem. Mabelle plans to express her protein fused to a polyhistidine-tag (His-tag). Explain the importance of His-tag in protein work.arrow_forwardA number of mutations affect the expression of the lac operon in E. coli. The genotypes of several E. coli strains are shown below. ("+" indicates a wild-type gene with normal function and "-" indicates a loss-of-function allele.) Please predict which of the following strains would have the highest beta-galactosidase enzyme activity, when grown in the lactose medium. CAP It P+ O`Z+ CAP+ I P+ O+ Zt CAP+ It P- O+ Z+ CAP+ It P+ 0+ Zarrow_forwardThe lac operon contains the genes that the bacterium needs to be able to use lactose as a carbon source. What would happen to the regulation of the Lac operon if: a) a mutation has occurred in the Lac Z gene which results in the formation of no functional β-galactosidase b) a mutation has occurred in the CAP protein so that cAMP cannot bindarrow_forward
- A transduction experiment was carried out to map the his and leu genes, which are involved in amino acid biosynthesis, of E. coli using a his+ leu+ donor and a his- leu- recipient. his+ leu- transductants will grow on agar media with: OA lacking histidine, containing leucine O B. containing histidine, containing leucine OC containing histidine, lacking leucine O D.a and b are correct O E. a, b and c are correctarrow_forwardAhmed had isolated bacteria from the sputum of a patient that seemed somewhat resistant to their antibiotic treatment, since multiple weeks of treatment had yielded no satisfactory improvement of their active disease. The researcher had then resuspended the patient's sputum in saline and used a mouse model to measure the efficiency of a phage selected for its capacity to target M. mycobacterium, as a single therapy, and as a combination therapy with antibiotics. Here is a schematic representation of the experiment: Group 1 Control (saline) Inoculate mice with patient sputum Group 2 wait for active Treated with phage disease to show Collect lung sample every 2 weeks Tuberculosis Sputum samples patient Group 3 Treated with Abx Group 4 Split mice showing active disease in 4 Treated with Abx and Count bacteria in sample groups phage Figure 4. Sputum from patients were isolated and resuspended in saline. Mice were given equal amounts of resuspended sputum intra-nasally and were left to…arrow_forwardAhmed had isolated bacteria from the sputum of a patient that seemed somewhat resistant to their antibiotic treatment, since multiple weeks of treatment had yielded no satisfactory improvement of their active disease. The researcher had then resuspended the patient's sputum in saline and used a mouse model to measure the efficiency of a phage selected for its capacity to target M. mycobacterium, as a single therapy, and as a combination therapy with antibiotics. Here is a schematic representation of the experiment: Group 1 Control (saline) Inoculate mice with patient sputum Group 2 wait for active disease to show Treated with phage Sputum samples Collect lung sample every 2 weeks Tuberculosis patient Group 3 Treated with Abx Split mice showing active disease in 4 Group 4 Treated with Abx and Count bacteria in sample groups phage Figure 4. Sputum from patients were isolated and resuspended in saline. Mice were given equal amounts of resuspended sputum intra-nasally and were left to…arrow_forward
- Antibiotics that target bacterial molecules not previously exploited are desperately needed. One such target is the protein FtsZ. The small molecule 3-methoxybenzamide (3-MBA) is known to inhibit FtsZ in Bacillus subtilis but is not bacteriocidal. Nonetheless, researchers reasoned that 3-MBA offered a good starting point for the synthesis of a molecule that might be a potential drug candidate. Over 500 3-MBA analogues were synthesized and screened; one called PA190723 was extremely potent in its capacity to bind FtsZ and inhibit bacterial growth. In fact, when used in a mouse model, PA190723 was bacteriocidal against methicillinand multidrug-resistant Staphylococcus aureus. What makes FtsZ a good drug target? What preliminary information about 3-MBA would be helpful if you were designing the 3-MBA analogues? As these researchers move forward with clinical (human) testing, what other parameters and outcomes must be assessed besides the bacteriocidal activity of PA190723?arrow_forwardwhat is the nature and likely location(s) of a mutant that would, 1)allow constitutive expression of the lac gene? 2)prevent the cell from responding to lactose ( genes are not induced when exposed to lactose)? 3) not allow the cell to utilize lactose even when the genes are inducedarrow_forwardVarious antimicrobial drugs to treat microbial infection have diverse mechanism of action. Consider the following antimicrobial drugs: A. Seconeolitsine, known as DNA topoisomerase I inhibitor in bacteria. (i) Explain briefly how inhibiting DNA topoisomerase I is a good mechanism of action for an antibiotic, include possible molecular machineries being targeted. (ii) What would be an appropriate response if seconeolitsine works well by stating the state of supercoiling in bacteria. (iii) To prove your answer (ii), you test the condition of bacterial DNA by running gel electrophoresis, one has been treated with seconeolitsine (+ sample) and the other one is not (- sample). Explain the position of each + sample and – sample band on the gel in reference to the point of origin (where you load your samples) or how far each DNA sample travel across agarose gel. (iv) Explain why you would expect answer (iii) for each + sample and – sample. B.…arrow_forward
- A number of mutations affect the expression of the lac operon in E. coli. The genotypes of several E. coli strains are shown below. ("+" indicates a wild-type gene with normal function and "-" indicates a loss-of-function allele.) Please predict which of the following strains would have the highest beta-galactosidase enzyme activity, when grown in the lactose medium. O CAP+ r* p* o* z O CAP* I P* o* z* O CAP* r* P O* z* O CAP I P* O z*arrow_forwardAhmed had isolated bacteria from the sputum of a patient that seemed somewhat resistant to their antibiotic treatment, since multiple weeks of treatment had yielded no satisfactory improvement of their active disease. The researcher had then resuspended the patient's sputum in saline and used a mouse model to measure the efficiency of a phage selected for its capacity to target M. mycobacterium, as a single therapy, and as a combination therapy with antibiotics. Here is a schematic representation of the experiment: Group 1 Control (saline) Inoculate mice with patient sputum Group 2 wait for active disease to show Treated with phage Sputum samples Collect lung sample every 2 weeks Tuberculosis patient Group 3 Treated with Abx Split mice showing active disease in 4 Group 4 Treated with Abx and Count bacteria in sample groups phage Figure 4. Sputum from patients were isolated and resuspended in saline. Mice were given equal amounts of resuspended sputum intra-nasally and were left to…arrow_forwardClary Leonhart used the pET vector system to express her prokaryotic amylase enzyme. She added peptone into her culture broth of BL21 (DE3) Escherichia coli strain. At the end of the experiment, she discovered that her protein was not expressed. She repeated three more times but her protein of interest was still not produced. (i) (ii) Explain the reason why Clary failed to obtain her protein of interest and suggest a solution to troubleshoot this problem. Clary plans to express her protein along with a polyhistidine-tag, or better known by its trademarked name IIis-tag. Explain the importance of His-tag in protein work.arrow_forward
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