Please explain it in long detail, then in a seperate answer summarize answer in
ID: 193224 • Letter: P
Question
Please explain it in long detail, then in a seperate answer summarize answer in 2 sentences or less.
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 vour 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
your experiment says that first of all you isolated the gene which is responsible for formation of bob protein and then you made a lot of copies of this gene by using PCR then after this process you inserted this DNA or gene into the plasmid vector.
Plamid is istransferred to the host cell for cloning.
The problem is that, all process is correct but protein is not formed in the host cell
This condition occurred due to various possible reasons:
1. Gene does not have necessary sequances or inappropriate sequances which are required for the formation of protein like promoter, coding region and terminator.
2. The polymerase not able to bind the Gene so transcription is affected.
3. Various other factors, proteins or enzymes are involved in the protein formation. Their presence or abscence affects the protein production.
4 presence of inhibitor or insufficient monomers like RNA nucleotides or amino acids availability.