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Biotechnology and Embryology Review Sheet i. DNA structure a. Packaging into chromosomes, Histones, nucleosomes b. Ss vs. ds DNA c. Introns & Exons (spliced mRNA & cDNA) ii. Bacteria as tools for Manipulating DNA & Gene regulation a. Know the 3 methods of utilizing bacteria to manipulate DNA. i. Transduction ii. Transformation iii. Conjugation b. What are Plasmids? – How are they utilized? c. Restriction enzymes – How do they work? i. Recombinant DNA & Sticky ends ii. DNA Ligase iii. Originate from Bacteria d. Reverse Transcriptase – Where was it first found? How is it utilized by scientists? i. cDNA – How is it different from regular DNA? ii. How are introns and exons relevant to cDNA iii. In GM bacteria-transformed, which DNA is used? e. Why are we interested in this type of DNA technology? i. What are some practical applications of recombinant DNA? ii. Historically what were the first organisms where this technology was used? iii. Genetically Modified Foods a. GM Crops and transgenic organism b. How can GM foods harm the environment or us? List examples. c. How can they benefit society? iv. Gene expression and regulation i. Prokaryotes vs. Eukaryotes ii. Differentiation and gene expression to form tissue and signals iii. Embryonic development 1. signaling 2. head-mRNA 3. homeotic genes-Homeoboxes 4. activators and repressor proteins 5. Promoter and operons v. DNA Technology a. Gel Electrophoresis i. How do you set up a gel? ii. What are some practical applications? iii. DNA Fingerprinting –There are two types. What is this technique used for? b. What are Probes and what are they used for? c. Restriction Enzymes and sticky ends d. PCR i. What is it? What is PCR used for? e. DNA Sequencing and the Human Genome Project (HGP) i. What are 3-4 benefits of the HGP? 1. Pharmacogenics – what is this? 2. Gene Therapy – what is this? ii. Why do we want to know the sequence of DNA? iii. What are some limitations? iv. How many genes do humans have? v. How many base pairs, how many genes and how many proteins f. Microarrays-genetic chips i. Used to study gene expression ii. Used to detect presence of certain genes in humans g. What are the ethical concerns? Explain the issues. vi. Stem cells & Cloning a. What is the difference between reproductive and therapeutic cloning? b. Where are the embryonic stem cells located in early embryonic development? c. Differences between dedifferentiated somatic cells, ESC and induced pluripotent stem cells? THERAPEUTIC: How does the immune system play a role in cloning? Rejection and Immuno-suppressants. d. Why are mammals harder to clone? e. What is nuclear transplantation and what is the benefit for stem cells vii. Embryology and Stem cells– Identify stages from drawings or figures (from lab or textbook) a. Fertilization i. Sperm – Form and Function 1. How is the sperm designed to penetrate the egg? a. Acrosomal reaction and fertilization envelope-function b. Where do sperms get energy to propel itself? ii. Egg – Form and Function 1. Why are eggs so large? What is the role of the vitelline membrane? b. Early Cleavage to Blastula– What is the purpose of this first phase of development? i. What do you start with and what do you end up with before cells differentiate? c. Gastrulation – What is the purpose of this second phase of development? i. 3 embryonic layers – What tissue will they developing into? 1. Ectoderm 2.Mesoderm 3. Endoderm ii. What are the first organs to form after gastrulation in most mammals? 1. Notochord 2. Neural plate – Neural folds – Neural Tube a. What does this develop into? d. Therapeutic cloning i. Stem cells. Know the various origins. ii. Steps to obtain ESC-nuclear transplant iii. Induced pluripotent stem cells