pET28a/VDH

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	pET28a/VDH was used to transformed E. coli BL21(DE3), and the resulting recombinant strain was used for VDH production. E. coli BL21(DE3) harboring pET28a/VDH was cultured in LB broth supplemented with 1% glucose and kanamycin (30 μL/mL) at 37 °C. The overnight culture was diluted 1:100 in fresh LB broth supplemented with kanamycin (30 μL/mL) and incubated at 30 °C until the optical density at 600 nm reached 0.8. VDH production was induced by adding 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG).

Protocol tips

pET28a/VDH was used to transformed E. coli BL21(DE3), and the resulting recombinant strain was used for VDH production. E. coli BL21(DE3) harboring pET28a/VDH was cultured in LB broth supplemented with 1% glucose and kanamycin (30 μL/mL) at 37 °C. The overnight culture was diluted 1:100 in fresh LB broth supplemented with kanamycin (30 μL/mL) and incubated at 30 °C until the optical density at 600 nm reached 0.8. VDH production was induced by adding 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG).

Publication protocol

After verifying the nucleotide sequence of vdh by sequencing, pET28a/VDH was used to transformed E. coli BL21(DE3), and the resulting recombinant strain was used for VDH production. E. coli BL21(DE3) harboring pET28a/VDH was cultured in LB broth supplemented with 1% glucose and kanamycin (30 μL/mL) at 37 °C. The overnight culture was diluted 1:100 in fresh LB broth supplemented with kanamycin (30 μL/mL) and incubated at 30 °C until the optical density at 600 nm reached 0.8. VDH production was induced by adding 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG), and the cultures were incubated overnight at 30 °C. E. coli BL21(DE3) harboring the empty vector pET-28a(+) was used as a negative control strain.

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