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| Proteins, Expression, Isolation and Analysis | The expression vector of pET-MaANS was confirmed and transformed into the E. coli strain BL-21. The E. coli strains carrying the recombinant plasmid were grown in LB medium and induced by 1-mM isopropyl β-d-thiogalactoside (IPTG) at 37°C for 2 h when the OD600 value reached 0.6. Cells were harvested by centrifugation and disrupted by sonication; the supernatant was collected and analyzed by SDS-PAGE with a 12% gel. High-affinity Ni-NTA Resin (GenScript, NJ, USA) was used for protein purification. | Get A Quote |
Background Flavonoids are a kind of important secondary metabolite and are commonly considered to provide protection to plants against stress and UV-B for a long time. Anthocyanidin synthase (ANS), which encodes a dioxygenase in the flavonoid pathway, catalyzes the conversion of leucoanthocyanidins to anthocyanidins, but there is no direct evidence indicating that it provides tolerance to stress in plants. Results To investigate whether ANS can increase tolerance to abiotic stress, MaANS was isolated from mulberry fruits and transformed into tobacco. Our results suggested that the bacterially expressed MaANS protein can convert dihydroquercetin to quercetin. Overexpression of MaANS remarkably increased the acc... More
