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Focus points chapters 6-9 for Exam #2 Chapter 6 1) Know the basic mechanism of bacterial cell division. Review how replication of the bacterial circular chromosome is accomplished. 2) Be able to label a growth curve with the 4 phase of bacterial growth and describe briefly what is happening at each in terms of metabolism. 3) Be able to describe 3 methods to estimate bacterial growth rate. 4) Be able to name 6 physical factors that limit bacterial growth. What are bacteria that that resist these limiting factors? (example cold tolerant = psychrophile). 5) What are sporulation and germination? What does this allow bacteria to do? Name 2 genus of spore formers. 6) What is the difference between selection, enrichment and, differentiation? Can you name a medium that does each of these? 7) What are the nutritional requirements for all bacteria. (ie how does a bug get Carbon etc.). Chapter 7 1) How does being haploid effect the susceptibility of bacteria to mutations? 2) Why do bacteria make great model organisms to study genetics? 3) How does DNA replication work in bacteria? 4) What is an operon? 5) Where does protein synthesis occur in bacteria? 6) Can you describe the “central dogma?” 7) What is feedback inhibition with regards to a biochemical pathway? Can this be administered at the genetic level? 8) Using the lac operon as a model system, describe gene regulation by de-repression. 9) Using the trp operon as an example, describe feedback inhibition. 10) What are spontaneous mutations? 11) How can mutations be induced? 12) What types of mutations are the most detrimental in terms of function of protein. (nonsense, missense, frameshift) 13) What is replica plating and how is it used to find mutations? 14) What is an auxotroph? What is a prototroph? 15) What is the Ame’s test? Since many carcinogens are also mutagens, is it always true that a mutagen is a carcinogen? How has the Ame’s test been used to fast track product safety? Chapter 8 1) Define the term “transformation” as it applies to bacteria. 2) When linear DNA is involved, transformation also involves homologous recombination. Why is this process less frequent than transformation using plasmids? 3) What is a plasmid and what are the features present on them. How have plamids been engineered to be useful to microbiologists? 4) How are antibiotic resistance genes on some plasmids used in genetic engineering? 5) What are some pharmaceutical products currently on the market that are produced by recombinant DNA? 6) Can you describe how a genetic fusion might be used to help manufacture and purify a protein? Go to the web and research “histag” gst- tag, Chapter 9 1) Describe how microbe organisms are named. 2) How can a dichotamous key be used to identify and organism? 3) Who are the members of the three domain classification system. 4) Why are archaebacteria closer to Eukaryotes than the Eubacteria they resemble? 5) What are some of the techniques used to make evolutionary phylogenies of bacteria? Study guide part deux for Exam #2 Chapters 6-9 After reading these chapters you should be able to answer the following questions What mechanism of fission do most bacteria use to reproduce? How do yeast reproduce? What are the 4 phases of bacterial growth? In which phase is bacterial growth the fastest? What are serial dilutions used for? What instrument is used to measure turbidity? Acidophiles like pH levels under ___ best? What is a mesophile with regards to temperature? What is an obligate anaerobe? obligate aerobe? microaerophile, facultative anaerobe. examples of these? What type of bacteria are extremely salt tolerant? Mycobacterium tuberculosis is described as a fastidious organism. Why? An enzyme that breaks down starch is called ______________ What types of bacteria make spores? What triggers sporulation? What is a plasmid? What kinds of things cause mutations? Which nucleotide base is only found in RNA? What is the product of transcription? What is “reverse transcription” of translation? How does it effect the central dogma? What enzyme is the most important in DNA replication? What does the enzyme ligase do? Why is ligase necessary for DNA replication? What is an Okazaki fragment? What direction does DNA polymerase synthesize DNA in? Which RNA contains the anticodon? Why is the genetic code considered degenerate? What does the term “feedback inhibition” refer to? Why are insertions of 3 bases into DNA less disruptive than insertions of one or two bases? What is a silent mutation? When bacteriophage transfer bacterial DNA instead of their own DNA, this is known as ______________. What is the “Ti” plasmid? What is a transposable element? Which restriction endonuclease cuts at the recognition sequence 5’ GAATTC 3’ ? RNA polymerase directs transcription of RNA from a DNA template. What sequence upstream of a gene attracts the polymerase? Who postulated the original 5 kingdom system? What is the proper way to write a bacterial name? A strain is a subgroup of what taxonomic classification? Why are the archaebacteria referred to as extremophiles? Why are ribosomes a good molecule to study taxonomy by molecular phylogeny? What is the “Ames” test? What is a “sticky end” ? Kary B Mullis owes his Nobel prize for PCR to the bacterium Thermus aquaticus do you think he shared the prize money? Make sure you review the posted essay questions for the exam as well. Essay questions for exam #2 Answer these questions in essay format to the best of your ability Essay question number one 1) Given your knowledge of the dual regulation of the lac operon, explain the observation that while the cell is growing on glucose in the medium, expression of B-galactosidase (lacZ) is low even if lactose is also present in the medium. You may use a diagram to accentuate your explanation, but be sure to explain the features of your diagram. Pretend that you are teaching the lac operon to a high school AP biology class. I will be looking for the following terms in your explanation: operator, promoter, repressor, positive activator, catabolite repression, and allostery. Essay question number two 2) Explain how the polymerase chain reaction (PCR) works and how this technique can be employed to: a) amplify a specific gene from a microorganism , and b) identify if a particular organism is present in a soil sample Describe the materials necessary to carry out a PCR reaction. You may draw a diagram, but your explanation should be able to stand alone. What are you going to add to the test tube?