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PBIO 427/527 Final Exam Review (These are not the only things to know, but they are key points.) What are Stanley Cohen and Herbert Boyer known for? What are enhancers and how do they work? Draw and label all of the regulatory and structural elements associated with a typical eukaryotic or prokaryotic gene. Describe what each of these elements does. Compare and contrast the following vector systems: plasmids, bacteriophage , YACs and BACs. What is a cDNA library and how is one made? What is a genomic library and how is one made? Explain how automated DNA sequence analysis and pyrosequencing is performed. Describe how the PCR is performed and outline its potential applications. How can a protein of interest be genetically engineered for production (and secretion) by E. coli? Why isn’t the plasmid that contains the strongest promoter always the best expression vector? Why are some recombinant proteins expressed in eukaryotic organisms (e.g., yeast, insects, plants, mice, sheep) rather than bacteria? Name five recombinant proteins approved for human use and name the corresponding disorders that they treat. Name and discuss the production of three non-protein commercial products produced in bacteria by genetic engineering. How does Bt toxin work and how is it genetically engineered for expression in transgenic plants? Compare and contrast the Ti plasmid method of plant transformation with the microprojectile bombardment method. Name 5 foreign genes that have been introduced and expressed in plants for the purpose of genetically engineering superior varieties of crop plants? Identify the foreign gene and the corresponding trait that it confers to the transgenic plant. Describe how transgenic animals are commonly produced and what kinds of tests are generally performed on the putative transgenic animals. What is a gene “knock-out” mouse and how would it be produced? Describe the two different strategies used to obtain the human genome sequence. Approximately how big is the human genome, how much of it codes for protein, how many genes are in it, and how many proteins are likely produced from it?? What is alternative splicing and how does this relate to the human genome project? What are microarrays and how can they be used to identify genes that are up-regulated or downregulated in human diseases. Name the two types of human gene therapy. What vectors are used to deliver this therapy? What is RNA interference (RNAi) technology? Explain how it works and what it does. Compare and contrast mammalian cloning versus therapeutic cloning. What is DNA fingerprinting? How is it done? What are the uses of this technology? What are patents and what criteria must they satisfy? Discuss the pros and cons of the human genome project. What principles should one keep in mind when considering an ethical issue? Considering these ethical principles, what is your view on an ethical issue (e.g., Bt-corn, ESCs)? Remember to review the various class presentations as questions will be written so test your knowledge of the major points in each instance.