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 conferring resistance to ampicillin and 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, and then added to bacteria growing in nutrient broth. 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 plus ampicillin, nutrient broth plus tetracycline, nutrient broth plus ampicillin and tetracycline, and nutrient broth without antibiotics. 7) Bacteria that contain the plasmid, but not the eukaryotic gene, would grow A) in the nutrient broth plus ampicillin, but not in the broth containing tetracycline. B) only in the broth containing both antibiotics. C) in the broth containing tetracycline, but not in the broth containing ampicillin. D) in all four types of broth. E) in the nutrient broth without antibiotics only. 8) Bacteria containing a plasmid into which the eukaryotic gene has integrated would grow in 9) Bacteria that do not take up any plasmids would grow on which media? 10) A principal problem with inserting an unmodified mammalian gene into a bacterial plasmid, and then getting that gene expressed in bacteria, is that 2
11) A gene that contains introns can be made shorter (but remain functional) for genetic engineering purposes by using A) RNA polymerase to transcribe the gene. B) a restriction enzyme to cut the gene into shorter pieces. C) reverse transcriptase to reconstruct the gene from its mRNA. D) DNA polymerase to reconstruct the gene from its polypeptide product. E) DNA ligase to put together fragments of the DNA that codes for a particular polypeptide.
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