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| Proteins, Expression, Isolation and Analysis | The bacteria were grown at 37 °C with shaking at 200 rpm in 500 mL of Luria-Bertani medium containing 100 mg L−1 ampicillin. After three hours, when the optical density at 600 nm (OD600) reached 0.6, protein expression was induced by adding 500 μL of 100 mM isotropy-β-D-thiogalactoside (IPTG), and the culture was incubated for approximately 4 h at 37 °C with shaking at 200 rpm. The protein was harvested by centrifugation of the culture and ultrasonication of the pellet at 4 °C. The recombinant protein was separated and purified using a high-affinity Ni-NTA resin column (L00250, Genscript Biotech) according to the instructions and then detected by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The eluted protein solution was further dialysed against 1 × PBS buffer for 12 h at 4 °C, which was repeated twice. The quality and concentration of the purified protein was determined using NanoDrop 2000, and the purified protein was stored at 4 °C. | Get A Quote |
Dehydrogenase ascorbic reductase (DHAR) is a critical enzyme involved in resistance to environmental stress. However, few DHAR genes have been identified in woody plants. Liriodendron chinense is a relict tree species of Magnoliaceae. Due to its outstanding ornamental features and fine wood properties, L. chinense is becoming increasingly applied in both timber production and landscaping. A broad ability to adapt is the initial premise of extensive planting of trees; as a result, discovery and functional validation of resistance genes are extremely important. In this study, a putative DHAR gene was isolated from L. chinense. The full-length coding sequence of the LcDHAR gene was cloned and expressed in ... More
