Study Guide for Biotechnology Proteins Quiz #2
Protein Translation, Antibodies, and Enzymes
TRANSLATION
-Describe eukaryotic protein production from DNA to RNA to protein. Be sure to include the role of promoters,
introns, exons, codons, tRNA, and peptidyl transferase.
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What are the steps in the process?
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Where does each step occur?
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What organelles are involved?
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What post-translational modifications could occur? Where does this happen?
-Compare and contrast eukaryotic and prokaryotic protein production.
-If given a sequence of DNA and the genetic code, determine the sequence of amino acids that would result.
-Compare and contrast point mutations and frameshift mutations. How does redundancy in the genetic code
affect mutations?
ANTIBODIES
-Describe the quaternary structure of an IgG antibody.
-Describe the function of an antibody using the terms antigen and epitope.
-What is the purpose/effect of antibody binding?
-Compare and contrast polyclonal antibodies with monoclonal antibodies.
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What are they?
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How are they produced?
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What are the pros and cons of each?
ELISA
-Proteins attach to the hydrophobic plastic.
-The purpose of the primary antibody is to bind to the protein of interest.
-The purpose of the secondary antibody is to bind to the primary antibody and produce a visible signal. The
secondary antibody is specific to the primary antibody and has an enzyme attached to it (or a fluorescent
marker). Addition of the enzyme’s substrate produces a visible color.
-What is the purpose of positive and negative controls?
-What would be the result of not washing properly after adding the primary antibody?
-What would be the result of improper washing after adding the secondary antibody?
-What are some practical applications of ELISA?
WESTERN BLOTTING
-Use the pre-lab questions as a study guide.
-The nitrocellulose membrane binds proteins. If blocker(milk protein) is not used to block the membrane, the
primary antibody will bind all over the membrane. Antibodies are proteins, too!
-What are some practical applications of Western Blotting?
ENZYMES
-Enzymes are proteins that speed up chemical reactions (catalysts).
-Describe the “Lock and Key” model using the terms substrate, active site, and products.
-Higher amounts of substrate allow for faster enzymatic turnover rates.
-Catalysts are unchanged by the reactions they catalyze, so they are available to continuously catalyze.
-Enzymes have an optimum temperature and pH at which they work fastest.
-Enzymes denature, at extreme temperatures and pH values, affecting the shape of the active site and
eliminating enzyme function. The temps and pHs at which this happens will vary depending on the enzyme.
Engineering enzymes to increase their active pH and temp ranges is a good way to make profits!
Know all vocabulary words on the Chapter 5(part two) lecture, excepting hybridoma and induced fit model.