Differential expression of the TwHMGS gene and its effect on triptolide biosynthesis in Tripterygium wilfordii

摘要: 3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) is the first committed enzyme in MVA pathway and involved biosynthesis of terpenes Tripterygium wilfordii. The full-length cDNA a 515 bp RNAi target fragment TwHMGS were ligated into pH7WG2D pK7GWIWG2D vectors to respectively overexpress silence, was overexpressed silenced T. wilfordii suspension cells using biolistic-gun mediated transformation, which resulted 2-fold increase drop 70% expression level compared with empty vector controls. During overexpression, TwHMGR, TwDXR TwTPS7v2 significantly upregulated control. In group, TwDXS, TwMCT visibly displayed downregulation produced twice higher than control value. These results proved that differential determined production triptolide laterally caused different trends relative gene terpene biosynthetic pathway. Finally, substrate acetyl-CoA docked active site TwHMGS, suggesting key residues including His247, Lys256 Arg296 undergo electrostatic or H-bond interactions acetyl-CoA.