20a - Biology, 8e (Campbell) Chapter 20 Biotechnology...

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Biology, 8e (Campbell) Chapter 20 Biotechnology Multiple-Choice Questions 1) If you discovered a bacterial cell that contained no restriction enzymes, which of the following would you expect to happen? D) The cell would be easily infected and lysed by bacteriophages. 2) Assume that you are trying to insert a gene into a plasmid. Someone gives you a preparation of genomic DNA that has been cut with restriction enzyme X. The gene you wish to insert has sites on both ends for cutting by restriction enzyme Y. You have a plasmid with a single site for Y, but not for X. Your strategy should be to C) Cut the DNA again with restriction enzyme Y & insert these fragments into the plasmid cut with the same enzyme. 3) What is the enzymatic function of restriction enzymes? D) To cleave nucleic acids at specific sites 4) How does a bacterial cell protect its own DNA from restriction enzymes? A) Adding methyl groups to adenines & cytosines 5) What is the most logical sequence of steps for splicing foreign DNA into a plasmid & inserting the plasmid into a bacterium? I. Transform bacteria with recombinant DNA molecule. II. Cut the plasmid DNA using restriction enzymes. III. Extract plasmid DNA from bacterial cells. IV. Hydrogen-bond the plasmid DNA to nonplasmid DNA fragments. V. Use ligase to seal plasmid DNA to nonplasmid DNA. C) III, II, IV, V, I 6) Bacteria containing recombinant plasmids are often identified by which process? C) Exposing the bacteria to an antibiotic that kills cells lacking the resistant plasmid Use the following information to answer the questions below. A eukaryotic gene has "sticky ends" produced by the restriction endonuclease EcoRI. The gene is added to a mixture containing EcoRI & a bacterial plasmid that carries two genes conferring resistance to ampicillin & tetracycline. The plasmid has one recognition site for EcoRI located in the tetracycline resistance gene. This mixture is incubated for several hours, exposed to DNA ligase, & then added to bacteria growing in nutrient broth. The bacteria are allowed to grow overnight & 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 plus ampicillin, nutrient broth plus tetracycline, nutrient broth plus ampicillin & tetracycline, & nutrient broth without antibiotics. 7) Bacteria that contain the plasmid, but not the eukaryotic gene, would grow D) In all four types of broth. 8) Bacteria containing a plasmid into which the eukaryotic gene has integrated would grow in E) The ampicillin broth & the nutrient broth. 1
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9) Why was the gene inserted in the plasmid before it was mixed with the bacteria? A) The plasmid acted as a vector to introduce the gene into the bacteria.
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This note was uploaded on 02/26/2011 for the course BIO 104 taught by Professor Smith during the Spring '11 term at Saddleback.

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20a - Biology, 8e (Campbell) Chapter 20 Biotechnology...

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