A eukaryotic gene has \"sticky ends\" produced by the restriction endonuclease E

Question

A eukaryotic gene has "sticky ends" produced by the restriction endonuclease EcoRI. The gene is added to a mixture containing EcoRI and a bacterial plasmid that carries two genes, which make it resistant to ampicillin and tetracycline. The plasmid has one recognition site for EcoRI located in the middle of the tetracycline resistance gene. The plasmid is cut by EcoRI, the eukaryotic gene is ligated in and the resulting plasmid with the gene inserted is transformed into bacteria. The bacteria are allowed to grow overnight and are streaked on a plate using a technique that produces isolated colonies that are clones of the original. Samples of these colonies are then grown in four different media: nutrient broth (liquid media with necessary nutrients only) plus ampicillin, nutrient broth plus tetracycline, nutrient broth plus ampicillin and tetracycline, and nutrient broth containing no antibiotics. The bacteria containing the engineered plasmid would grow in... Othe ampicillin and tetracycline broth only the nutrient broth, the ampicillin broth, and the tetracycline broth. the nutrient broth and the tetracycline broth only. the nutrient broth only. the ampicillin broth and the nutrient broth.

Explanation / Answer

The recognition site is located in the middle of tetracycline resistance gene.

Since the plasmid is cleaved at the middle of tetracycline resistance gene, the resistance for tetracycline is lost.

Therefore, the genetically engineered plasmid will contain only ampicillin resistant gene.

The bacteria containing the genetically engineered plasmid would grow in both nutrient broth and the ampicillin broth.

Therefore, option E is correct.

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