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How does the protein product (beta-lactamase) of the Amp-R gene confer resistance to the antibiotic ampicillin? Find the specific mechanism of ampicillin resistance for bacteria expressing beta-lacamase via pUC19.

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Patrina Schowalter
Patrina SchowalterLv2
29 Sep 2019
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Related questions

The introductory passage explains that Vibrio cholerae bacteria may benefit from being able to obtain genes from their prey. What would be the best way to test whether they can benefit within one week of exposure to prey with a gene that confers resistance to ampicillin, an antibiotic that is commonly used in microbiology laboratories? Choose all of the steps that you should include.

Transfer V. cholerae that have been exposed for one week to prey with the resistance gene to medium with ampicillin.
Transfer V. cholerae that have been exposed for one week to prey lacking the resistance gene to medium with ampicillin.
Set up V. cholerae cultures that are resistant to ampicillin and allow them to grow for a week in the presence of prey with the resistance gene on medium that does not contain ampicillin.
Set up V. cholerae cultures that are susceptible to ampicillin and allow them to grow for a week in the presence of prey that have the resistance gene on medium that does not contain ampicillin.
Set up V. cholerae cultures that are susceptible to ampicillin and allow them to grow for a week in the presence of prey lacking the resistance gene on medium that does not contain ampicillin.
Set up V. cholerae cultures that are susceptible to ampicillin and allow them to grow for a week in the presence of prey lacking the resistance gene on medium that contains ampicillin.
Set up V. cholerae cultures that are susceptible to ampicillin and allow them to grow for a week in the presence of prey that have the resistance gene on medium that contains ampicillin.
Set up V. cholerae cultures that are resistant to ampicillin and allow them to grow for a week in the presence of prey lacking the resistance gene on medium that does not contain ampicillin.

Recognize the following major structures and know how changing the structure will effect the activity (if we discuss any):

Penicillins, Cephalosporins

1. What is the mechanism of action for penicillins and cephalosporins? Is it bactericidal or bacteriostatic? What are the side effects?

2. What is the common structural feature of all penicillins and which is the functional group that is modified?

3. What are the common problems associated with natural pencillins?

4. What does hypersensitivity have to do with chemical reactivity?

5. What is the major mechanism for resistance with the penicillins? List two other mechanisms.

6. What is the structural change that we make to instill beta lactamase resistance? This reduces the activity of these penicillins, what are the three undesirable characteristics. Consider nafcillin as the prototypes. What advantage do oxacillin & dicloxacillin have over nafcillin?

7. What is the normal spectrum of susceptible organisms for pen V, limited (methicillin), broad (ampicillin) and extended (carbenicillin) spectrum penicillins?

Class/Spectrum

Gram (+) coverage

Gram (-) coverage

Beta-lactamase resistant

other

Pen V/traditional

+

- (weak)

-

Methacillin-like/limited or narrow

Carbenicillin/extended

Inc. pseudomonas

Ampicillin/Broad

A plasmid that carries two antibiotic resistance genes, to ampicillin and tetracycline, respectively, has one restriction site for EcoRI located within the tetracycline resistance gene. Eukaryotic DNA containing a gene of interest (g.o.i.) has been digested with diffferent restriction enzymes, none of which cut within the gene of interest sequence). The resultant fragments are added to plasmid DNA that has been digested with EcoR1 only; the mixture was then incubated with DNA ligase. Resulting plasmids (recombinant and otherwise) are added to competent bacteria lacking resistance genes and growing in nutrient broth. The bacteria are then plated and allowed to grow overnight producing isolated colonies. Samples of these colonies are then grown in four different media: nutrient broth plus ampicillin; nutrient broth plus tetracycline; nutrient broth plus ampicillin and tetracycline; and nutrient broth containing no antibiotics.

1. The bacteria containing the plasmid with the gene of interest successfully insertedin would grow in

A. Tthe nutrient broth only.

B. The nutrient broth, the ampicillin broth, and the tetracycline broth.

C. Tthe nutrient broth and the tetracycline broth only.

D. The ampicillin broth and the nutrient broth.E.Tthe ampicillin and tetracycline broth only.

2. Bacteria transformed with the plasmid alone (not containing any eukaryotic DNA), would grow

A. Only in the broth containing both antibiotics.

B. In the nutrient broth plus ampicillin, but not in the broth containing tetracycline.

C. In the broth containing tetracycline, but not in the broth containing ampicillin.

D. Only in the broth that contained no antibiotics.

E. In all four types of broth.

4. Bacteria that did not take up any plasmids at all would grow in which media?

A. The tetracycline and ampicillin broth only

B. The nutrient broth and the tetracycline broth only

C. The nutrient broth and the ampicillin broth only

D. The nutrient broth only

E. All four broths

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