I wish to express human NAT10 enzyme in a bacterial system. Are there any codon optimized commercially available bacterial vector plasmids that facilitate this? If this is not feasible then what kinds of experimental approaches would be beneficial for expressing human NAT10? Appreciate all of your suggestions. Thanks
Expressing human NAT10 in E. coli can be challenging due to its size, eukaryotic origin, and potential toxicity. Codon-optimized NAT10 gene synthesis (e.g., via GenScript, IDT) cloned into bacterial expression vectors like pET28a(+) or pGEX-6P-1 may improve success. Use strains like Rosetta(DE3) or BL21(DE3) pLysS to enhance expression of rare codons. Test lower induction temperatures (16–18°C) and solubility tags (MBP, GST) to enhance folding. If insoluble, consider eukaryotic systems (baculovirus in Sf9 or HEK293) for functional protein.
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