| Products/Services Used | Details | Operation |
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| Proteins, Expression, Isolation and Analysis | Target proteins were individually eluted with elution buffer (20 mM maltose in binding buffer), and 1-mL fractions were collected. For each GES homolog, 15-μL aliquots of every other fraction were analyzed using SDS-PAGE with gradient (10% acrylamide) gels, and proteins were visualized through an eStain L1 protein staining system (Genscript, USA). Fractions containing each MBP-tagged GES (molecular mass of approximately 109 kDa) were combined and dialyzed in PD-10 desalting columns (GE Healthcare) following the manufacturer’s instructions. For the removal of the MBP tag, Factor Xa (BioLabs, China) was used to cut the MBP tag in buffer (20 mM HEPES, 500 mM NaCl, 2 mM CaCL2, and 50% glycerol, pH 8.0). The reaction mixture was incubated at 23 °C for 6 h. The protein concentration was determined using the Bradford method, with bovine serum albumin used as the standard. | Get A Quote |
Iridoid compounds have been reported to be accumulated in the dried fruits of Gardenia jasminoides (Rubiaceae), which are used in traditional Chinese medicine. These compounds exhibit obvious pharmacological activities, including effective protective effects on the liver and therapeutic efficacy for cardiovascular diseases. However, the biosynthetic pathway of iridoid remains uninvestigated. In this study, 12 transcriptomes from four tissues (lower leaves, top leaves, flowers, and fruits) were analyzed to characterize the accumulation of active constituents and to identify the corresponding genes involved in iridoid biosynthesis. Almost all genes in the mevalonate (MVA) pathway and the 2-C-methyl-d-erythritol... More
