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Chapter 18-21 1. The restriction enzyme, HindIII recognizes the sequence 5’-AAGCTT-3’, cutting between the two A’s. Draw the double-stranded sequence before and after the enzyme cuts. What type of bonds are being cleaved by the restriction enzyme? 2. One strand of a DNA molecule has the sequence: 5’-CCTTGACGATCGTTACCG-3’. Draw the other strand. The restriction enzyme PvuI recognizes the sequence 5’-CGATCG-3’. Will this enzyme cut this DNA molecule? If so then please indicate where. 3. Describe in detail how to clone a human gene in a bacterial plasmid. 4. Describe in detail how to amplify DNA by using PCR. 5. Describe the role of complementary base pairing during Southern blotting, DNA sequencing, northern blotting, RT-PCR, and microarray analysis. 6. Describe how some biotechniques can be used to diagnose diseases. 7. Describe how scientists can bioengineer viruses to deliver proper genes to tissues that have mutated genes. 8. Describe how plants are bioengineered? 9. Describe some of the concerns regarding safety and ethics with bioengineering. Ok Guys.... the next set of questions 10-13 we will do activities over next week... so look at them if you don't understand.... you will have to come back to them..... :) 10. The following is a single strand of DNA. Find all potential “6-base cutter” restriction sites that would exist in the actual double stranded DNA molecule. 5’-TAGAATTCGACGGGATCCGGGGGGCATGCAGATCA-3’ 11. A bacterial plasmid is 100 kb in length. The plasmid DNA was digested to completion with two restriction enzymes in three separate treatments: EcoRI, HaeIII, and EcoRI + HaeIII (double digest). The fragments were then separated with electrophoresis, as shown. a. Using the circle provided, construct a labeled diagram of the restriction map of the plasmid. Explain how you developed your map. b. Describe how: i. Recombinant DNA technology could be used to insert a gene of interest into a bacterium c. ii. Recombinant bacteria could be identified iii. Expression of the gene of interest could be ensured Discuss how a specific genetically modified organism might provide a benefit for humans and at the same time pose a threat to a population or ecosystem. 12. Many human genes contain introns. Since bacteria cannot excise introns from mRNA, explain how bacteria can be used to make large quantities of a human protein. 13. A pregnant 30-yr-old woman learns her father has Parkinson’s disease. There is no history of the disease in her husband. A genetic test that can detect the disease is used on her father, husband, fetus, & on her. Autoradiographic results after Southern blotting are shown below. Father ___ Husband ___ Woman ___ Fetus ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ Will the woman develop the disease? Explain. Will the child develop the disease? Explain. 14. As you learned in mitosis, the parent cell gives rise to two daughter cells that are genetically identical to the parent cell. Yet you, the product of many mitotic cell divisions are not composed of identical cells. Why? 15. During development, the lower cells are synthesizing signaling molecules, where as the upper cells are expressing receptors for those molecules. In terms of gene regulation, explain how these cells came to synthesize different molecules. 16. The DNA sequences called homeoboxes, which help homeotic genes in animals direct development, are common to flies and mice. Given this similarity, explain why these animals are so different.