Hello,
I am currently working on making a fusion protein through the tagging of an endogenous gene. I have restricted and ligated fragments such there has appeared a 3 amino acid (Alanine repeats) insertion after the stop codon of my tag protein and before the 3' UTR start of the endogenous protein. This has not resulted in any frame shifts (The endogenous protein, tagged protein and UTRs are in frame) but I am curious to know if these 3 amino acids might affect the folding or expression of the fusion protein since they were not inherently present in the endogenous 3' UTR sequence.
Having a few amino acids in the spacer region between two proteins are not likely to impact folding or expression, since the two protein domains are most likely going to fold independently. However as Sebastian Schmitt
pointed out, if you have a a stop codon in your upstream tag protein then you are not going to generate any fusion protein.- Badges
- Science topic