Part:BBa_K4726000
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.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Unknown
- 1000COMPATIBLE WITH RFC[1000]
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.
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