1. You have cloned the gene sequence of the novel eukaryotic protein you have na
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Question
1. You have cloned the gene sequence of the novel eukaryotic protein you have named Protein Bob (2,100 bp in length). You have used PCR to amplify up the Protein Bob gene from your isolated genomic DNA. After growing up colonies and testing the plasmid you discovered that your cloning protocol has gone according to plan and you did every step correctly, there was no human error. Yet, there is no functional protein production. Explain the reasoning behind this problem. How would you rectify the problem? (10 Points)Explanation / Answer
the reason behind this problem might be that:
Eukaryotic genes have introns and exons, interpersed with each other. Before translation, the pre-mRNA is spliced to remove the introns. This splicing machinery is not present in the prokaryotic cells (bacterial cells where the protein is tried to expressed). The genomic DNA would have the entire gene of protein Bob along with its introns which is pehaps disordering the translation step.
This problem can be rectified by using the cDNA of the protein bob, to make the clone.
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