DNA Replication and Repair
Textbook Readings:
Allison, Chapters 6 and 7
Questions:
Question 1.
You’ve discovered a novel protein and suspect that it is a DNA topoisomerase. Describe how you will design an in vitro experiment to test for the activity of this novel protein to demonstrate that it is a DNA topoisomerase.
Question 2.
There’s a new mammalian virus you’re studying. It has a 200 kb double-stranded DNA genome. Because of its large genome size, you think it may have more than one origin of replication. Explain an experiment to map the origins of replication in this new mammalian virus.
Question 3.
Explain what the effect would be on the reading frame and gene function if three nucleotides were inserted into the middle of an mRNA? Would a frameshift occur? Why or why not?
Question 4.
Explain what the effect(s) would be on the reading frame and gene function after exposure to an alkylating agent which induces the formation of O6-methylguanine . What effects would there be after DNA replication?
References
Recombinant DNA, Cloning and Tools for Analysis of Gene Expression
Textbook Readings:
Allison, Chapters 8 and 9
Questions:
Question 5.
Below are the first fifty nucleotides in the sequence of the mouse fetal beta globin epsilon gene. You are interested in how the fetal beta globin epsilon gene is expressed; spatially, temporally and/or constitutively. Design and describe an experiment making use of the provided nucleotide sequence to analyze fetal beta globin epsilon gene expression. (Sequence provided is the nucleotide sense strand from Genbank).
5’-gacaacctca agtctgcctt ggccaagctc agtgaactgc actgtgacaa-3’
Question 6.
You’ve been provided with the complete nucleotide sequence for the domestic dog pancreatic amylase AMY2B gene. Explain how you would search for and locate a homologous gene in the Fennec fox, assuming it has had its genome sequenced. Describe how you would obtain the original amylase protein, from each test specimen without obtaining it from the dog or Fennec fox directly (no blood, tissue or serum draws), and test it for comparative activity between the domestic dog and the Fennec fox. (Note, no field work or hands-on work with live dogs or Fennec foxes).
Question 7.
You’re working on a newly discovered gene and are tasked to quantitatively quantify the precise transcript levels of this gene being expressed in a culture of cells. Explain the method you will use and details of how you will perform this.
Question 8.
You have purified a novel protein and have a small aliquot of it. Unfortunately, there’s no commercially available antibodies to this new novel protein of yours. What can you do to obtain monoclonal antibodies to your new, novel protein so that you can use an antibody to screen sets of tissue samples to determine if your protein is expressed? You also want to have antibodies to use for a later time for other possible experiments.
Question 9.
Identify a country or government agency that has a published policy focused on the regulation of genetically modified organisms.
Question 10.
Once you have identified your country of focus, summarize their main stance on the regulation of biotechnology products, especially GMOs. Are GMOs banned in any way or form?
References