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Unformatted text preview: APPENDIX 4 for lactose utilization would not be made, whether or not lac- tose was present. b. The lac genes would continue to be transcribed and the enzymes made, whether or not lactose was present. c. Same predicted result as for b. d. RNA polymerase would not be able to transcribe the genes; no proteins would be made, whether or not lactose was present. use it as a template to make cDNA (using reverse transcriptase). Cloning this DNA rather than the entire genome would yield a smaller library of genes to be screenedionly those active in liver cells. Furthermore, the genes would lack introns, making the desired gene easier to manipulate after isolation. 10. c (Bacteria lack the RNAesplicing machinery needed to delete eukaryotic introns.) 10- White, reflecting the genetic makeup 0f the nucleus donor 11. Isolate plasmids from a culture of E. coli. Cut the plasmids and 11. A mutation in a single gene can influence the actions of many other genes if the mutated gene is a control gene, such as a home, otic gene. A single control gene may encode a protein that affects (activates or represses) the expression of a number of other genes. In addition, some of the affected genes may themselves be control genes that in turn affect other batteries of genes. Cascades of gene expression are common in embryonic development. 12. The protein to which dioxin binds in the cell is probably a tran7 scription factor that regulates multiple genes (see Module 11.3). If the binding of dioxin influences the activity of this transcrip tion factorieither activating or inactivating itidioxin could thereby affect multiple genes and thus have a variety of effects on the body. The differing effects in different animals might be explained by differing genetic details in the different species. It would be extremely difficult to demonstrate conclusively that dioxin exposure was the cause of illness in a particular individ7 ual, even if dioxin had been shown to be present in the person's tissues. However, if you had detailed information about how dioxin affects patterns of gene expression in humans and were able to show dioxin-specific abnormal patterns in the patient (perhaps using DNA microarrays; see Module 11.9), you might be able to establish a strong link between dioxin and the illness. 13. Wilmut was able to coordinate the cell cycle of the donor and host cells by depriving them of nutrients. When faced with starvation, both cells switched to the G0 phase of the cell cycle. Wilmut could therefore be sure that, when placed in a growth medium with nutrients, the cell cycles within both the donor and host cell were synchronized. This allowed him to success frilly clone a mammal from an adult cell for the first time. Chapter 12 1. a. PCR; b. arestriction enzyme; c. gel electrophoresis: d. nucleic acid probe; e. cloning 2.d3.b4.b5.c5.c 7. Because it would be too expensive and time consuming to com, pare whole genomes. By choosing STR sites that vary consider, ably from person to person, investigators can get the necessary degree of specificity without sequencing the entire genome. 8. Medicine: Genes can be used to produce transgenic lab animals for AIDS research or for research related to human gene therapy. Proteins can be hormones, enzymes, blood—clotting factors, or the active ingredient of vaccines. Agriculture: Foreign genes can be inserted into plant cells or animal eggs to produce trans, genic crop plants or farm animals. Animal growth hormones are examples of agriculturally useful proteins that can be made using recombinant DNA technology. 9. She could start with DNA isolated from liver cells (the entire genome) and carry out the procedure outlined in Module 12.1 to produce a collection of recombinant bacterial clones, each car, rying a small piece of liver cell DNA. To find the clone with the desired gene, she could then make a probe of radioactive RNA with a nucleotide sequence complementary to part of the gene: GACCUGACUGU. This probe would bind to the gene, labeling it and identifying the clone that carries it. Alternatively, the biochemist could start with mRNA isolated from liver cells and A—11 Chapter Revlew Answers the human DNA containing the HGI—l gene with the restriction enzyme to produce molecules with sticky ends. Join the plasi mids and the fragments of human DNA with ligase. Allow E. coli to take up recombinant plasmids. Bacteria will then replicate plasmids and multiply, producing clones of bacterial cells. Identify a clone carrying and expressing the HGl-I gene using a nucleic acid probe. Grow large amounts of the bacteria, and extract and purify HGI—l from the culture. 12. Determining the nucleotide sequences is just the first step. Once researchers have written out the DNA ”book,” they will have to try to figure out what it meansiwhat the nucleotide sequences code for and how they work. 13. Some issues and questions to consider: What are some of the unknowns in recombinant DNA experiments? Do we know enough to anticipate and deal with possible unforeseen and negative consequences? Do we want this kind of power over evolution? Who should make these decisions? If scientists doing the research were to make the decisions about guidelines, what factors might shape their judgment? What might shape the judgment of business executives in the decisionemaking pro cess? Does the public have a right to a voice in the direction of scientific research? Does the public know enough about biology to get involved in this decisionemaking process? Who represents ”the public," anyway? 14. Some issues and questions to consider: What kinds ofimpact will gene therapy have on the individuals who are treated? On society? Who will decide what patients and diseases will be treated? What costs will be involved, and who will pay them? How do we draw the line between treating disorders and "improving" the human species? 15. Some issues and questions to consider: Should genetic testing be mandatory or voluntary? Under what circumstances? Why might employers and insurance companies be interested in genetic data? Because genetic characteristics differ among ethnic groups and between the sexes, might such information be used to dis- criminate? Which of these questions do you think is most impor tant? Which issues are likely to be the most serious in the future? 16. Gather two groups of volunteers. Have one group ingest a stand- ard amount of GMO corn in their diet and the other group in- gest a standard amount of traditional corn. Monitor the health of the individuals in both groups, looking for differences. In real life, such a study would be problematic because it is difficult to control and monitor what people eat, it would require many years to search for long—term health effects, and it may run afoul of ethics standards for human testing. Chapter 13 1. According to Darwin’s theory of descent with modification, all life has descended from a common ancestral form as a result of natural selection. Individuals in a population have hereditary variations. The overproduction of offspring in the face of limited resources leads to a struggle for existence. Individuals that are well suited to their environment tend to leave more offspring than other individuals, leading to the gradual accumulation of adaptations to the local environment in the population. ...
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