Worksheet Questions 5

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Biology

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Jan 9, 2024

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1 WORKSHEET QUESTIONS 5 (Total: 6 marks) Due October 25 by 2 pm EST 1. An extract of developing Drosophila melanogaster embryos was loaded onto a two-dimensional gel and subsequently, the polypeptides were transferred onto a nitrocellulose membrane for Western Blot analysis. Anti-UBX monoclonal antibodies were used to detect the HOX protein, UBX which is essential for embryonic development of D. melanogaster . The Western Blot result shown below shows multiple signals/spots, each with decreasing molecular weight from low pH to high pH region (spots 6 to 1). Answer the questions below based on the above information. a) Why multiple signals/spots are observed for the same protein? (2 marks) This would mean that there are multiple proteins with similar if not same molecular weight. This distorts the data since we don’t know the actual amount of protein detected and can only compare ones we see. b) Why do the signals/spots have a certain pattern, i.e. , increasing in molecular weight as we go towards the low pH region (from spots 1 to 6)? How can you explain this pattern? (2 marks) Lower pH means a higher negative charge, and since SDS works based off making all the proteins negative so it can move towards the bottom of the gel in one direction it would be noticed that the higher the negativity the stronger it is.

2 c) How can you prove or disprove that the spot pattern is due to the existence of differentially phosphorylated UBX isoforms? (2 marks) You would leave out the HOX protein which the protein UBX binds to. If you do this and end up with no result, you therefore prove that the spot pattern is due to UBX binding to the HOX protein.

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Worksheet Questions 5

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