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BIO SCI 98 Study Guide - Midterm Guide: Two-Dimensional Gel Electrophoresis, Isocitrate Dehydrogenase, Pyruvate Dehydrogenase

Biological Sciences
Course Code
Pavan Kadandale
Study Guide

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Bio98 W2018 Practice Midterm 3
01. I inject the muscles of wildtype and mutant mice with radiolabeled glucose. I then
measure the amount of radiolabeled lactate that is present in the blood before, during
and after exercise. My data is shown in the graph.
02. I am attempting to create the Glycolytic pathway (GlcPyruvate) in vitro. I purify all the
enzymes of the pathway, the co-factors required, NAD+, ADP, ATP and Pi. I then mix all these
components together and add Glucose. I then measure the ratio of Pyruvate:Glucose in the
mixture over time. My data is shown in the graph. At the point marked “X” in the graph, I
added a non-protein substance to the pathway.
03. In order to make DNA for replication, a cell must use up some TCA intermediates, which would decrease the TCA. But
making DNA also requires energy. So how does the cell overcome this paradox of needing energy, but taking away
intermediates from the cycle that provides energy?
04. A specific mutation in the promoter of the gene that encodes the enzyme Isocitrate dehydrogenase causes the
promoter to become weaker, and for transcription of the protein to decrease. I take equal numbers of wild type cells
and cells that are heterozygous for the mutation, and measure the kinetics of Isocitrate dehydrogenase in the cells.
05. A lack of oxygen causes cells to convert pyruvate to lactate to regenerate the NAD+ required for Glycolysis.
06. I perform a 2D gel electrophoresis of proteins from the eggs of two different
species of birds. I then immunoprecipitate the protein marked “X” in species 1
using an antibody specific to that protein. The same antibody
immunoprecipitates the protein marked “Y” in species 2. After treating the
purified proteins with Glycosidase, they run identically on the 2D gel.
Sequencing shows that X and Y have identical amino acid sequences.
07. I am studying a newly discovered virus. I find that the virus causes infected cells to die. I now
take cells and expose them to either the virus, or virus treated with Glycosidase and after 3
days, measure the number of live cells present. My data is shown (the number of infection-
capable viruses is the same before and after Glycosidase treatment).
08. Mark each statement below TRUE or FALSE and give reasons. You will get credit ONLY if your reasoning is correct.
a) Gluconeogenesis is upregulated during prolonged exercise since blood sugar levels drop
b) A substrate cycle between DHAP and GA3-P might be used by some organisms to generate heat
c) A mutation in Adenylate Cyclase would most likely decrease the rate of Gluconeogenesis in liver
09. Imagine that I start off with 10 molecules of oxaloacetate in a mitochondria. I now allow the TCA to run just enough to
generate 20 molecules of CO2 (assuming that oxaloacetate is being used only for TCA in these mitochondria).
10. I have mutant cells that make no PFK2 enzyme. I measure rates of oxygen consumption in these mutant cells and
compare them to wild type cells.
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