Difference between revisions of "Part:BBa K4726000"

Revision as of 00:12, 10 October 2023

N-acetyl-α-D-galactosamine deacetylase_Flavonifractor plautii

This encodes a N-acetyl-α-D-galactosamine deacetylase from Flavonifractor plautii originating from the human gut microbiota.

This enzyme removes the acetyl moiety of N-acetyl-α-D-galactosamine (A antigen). This however does not results in full degradation of A antigen a α-D-galactosamine residue remains.

The expression of this enzyme led to significant proteotoxicity shown by an impaired growth of Escherichia coli BL21 (DE3). We thus recommended to use Escherichia coli Rosetta (DE3) pLysS which helps stabilizing pET recombinants encoding proteins.

To clone this genetic construct onto our expression vector (pET-3a) we added NdeI site in 5' and BamHI site in 3' of the gene. We added TATATA sequences before NdeI site and after BamHI site to help with the restriction digestion.