3.We anticipate our recombinant DHFR protein to be functional for use in future research applications. However, if you had been able to test functionality and found that your recombinant GST-DHFR-His protein was not functional, what would you propose is one potential issue you would investigate to troubleshoot further. In other words, what is one thing that could have gone wrong to result in non- functional protein from the method used and results obtained this semester

In the given case, the protein would have required to go through the process of post-translational modification, however, the mentioned protein would have been cultured in the prokaryotic system, the process of post-translational modification would have failed to take place. This step would be essential for the functionality of the protein. Thus, there is a need to culture it within the eukaryotic system.

The covalent and generally the enzymatic modification of proteins post its biosynthesis is termed as post-translational modification.