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Table 3 Primers for expression, disruption, and complementation of the hno gene

From: Green route to synthesis of valuable chemical 6-hydroxynicotine from nicotine in tobacco wastes using genetically engineered Agrobacterium tumefaciens S33

Primers

Sequence (5′–3′)

hno-F

CGGGATCCGATGACAGAAAAGATATATGATGC (BamHI)a

hno-R

CCCAAGCTTTTAAGCGGTCGCCTTC (HindIII)a

Primer A

CGCGGATCCATGACAGAAAAGATATATGATGC

Primer B

TCGATAAATGCGCGCTTTTAGGCTAGCTGCAACTCGTTC

Primer C

AAAAGCGCGCATTTATCGAACACGCCGAGATGGCTGAC

Primer D

GCCGGGCCCTTAAGCGGTCGCCTTC

Primer F2

GCCGGGCCCCATGACAGAAAAGATATATGATGC (ApaI)a

Primer R2

CGCGGATCCTTAAGCGGTCGCCTTC (BamHI)a

  1. aThe restriction site is underlined