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Exam III / Final Exam Study Guide Chapter 18 Know the biology of the Ti plasmid and the binary Ti plasmid system Explain how the binary Ti plasmid system and the microprojectile bombardment (biolistic) system is used to genetically engineer plants (i.e., know how to genetically engineer plants for expression of your favorite gene) What are reporter genes and how do they work? Chapters 19 and 20 Know what genes have been genetically engineered in plants and what new traits they confer as discussed in class Be able to explain how to modify any given gene/cDNA for expression in plants (i.e., gene construction) and how to introduce it into plants Know how to test plants for transgene insertion, gene expression, and protein production Know the reasons for non-identical expression of foreign genes in genetically modified plants Chapter 21 Understand the use of microinjection and engineered stem cells in creating transgenic animals Understand how to make and select genetically engineered stem cells for gene knockouts Know the Cre-loxP system for gene knockouts Know the reasons for non-identical expression of foreign genes in genetically modified organisms (GMOs) Be familiar with the animal genetic engineering examples covered in class Be able to explain mammalian cloning via nuclear transfer Chapters 22 Understand how biotechnology is regulated Chapters 23 Recognize the ethical issues resulting from biotechnology Be able to express your personal views on this issues based on scientific information Sample essay questions 1. Explain how you would go about creating a genetically engineered goat that expresses human growth hormone in its milk? 2. Humans are now eating food from genetically modified organisms (GMOs), particularly from plants. Give five examples in which you identify the genetically engineered plant, the altered trait, and the gene construction responsible for this trait. List any concerns that opponents have expressed with regard to the specific examples you have selected. 3. Explain how you would produce a clone of yourself or a favorite mammalian pet. The items below represent many key points from this course that you should know. Structural organization of prokaryotic and eukaryotic genes Transcription and transcriptional regulation in prokaryotes and eukaryotes Enhancers and how they work Restriction enzymes, DNA cloning, and cloning vectors cDNA & genomic library production & screening; characterization of clones Southern, northern, and western blotting PCR methodology and applications (Reverse transcriptase-PCR, Real time-PCR) Automated and manual methods of DNA sequencing, including pyrosequencing Monoclonal antibody production and ELISAs DNA fingerprinting (especially VNTRs and STRs) Genetic engineering/expression of eukaryotic genes in prokaryotes and vice versa Protein targeting sequences and their addition to genetically engineered genes RNAi and other methods to eliminate specific RNAs Human gene therapy (basic types, vectors involved, some examples) Application of recombinant DNA technology to vaccine production (e.g., subunit vaccines) Use of the binary Ti plasmid system and microparticle bombardment (biolistic method) in plant genetic engineering Be familiar with the plant genetic engineering examples covered in class Use of microinjection and engineered stem cells in creating transgenic animals Making and selecting genetically engineered stem cells for gene knockouts Cre-loxP system for gene knockouts Reasons for non-identical expression of foreign genes in genetically modified organisms (GMOs) Be familiar with the animal genetic engineering examples covered in class Mammalian cloning via nuclear transfer Key points of the Human Genome Project (including human genome size, the two approaches used to get the human genome sequence and ethical implications) Be able to understand and design a microarray experiment Know key points of the student presentations to answer multiple choice questions