EC Number   |
|---|
   5.5.1.12 | DNA and amino acid sequence determination and analysis of 18 different amino acid sequence of copalyl diphosphate synthase, CPS, phylogenetic analysis and tree |
| 5.5.1.12 | expression in Escherichia coli |
| 5.5.1.12 | expression of His-tagged enzyme in Escherichia coli |
| 5.5.1.12 | functional expression of Picea abies isopimara-7,15-diene synthase (diTPS-ISO) in Nicotiana benthamiana leaves via Agrobacterium tumefaciens transfection method, coexpression with a multifunctional cytochrome P450 (CYP720B4) from Picea sitchensis, subcloning in Escherichia coli strain NEB10beta |
| 5.5.1.12 | gene CPS1, phylogenetic analysis and tree, functional recombinant expression in Saccharomyces cerevisiae strain AM113, with co-overexpressing of GGDPS1 from Cistus creticus, and Nicotiana benthamiana, heterologous expression of the RoCPS1 protein in Escherichia coli fails |
| 5.5.1.12 | gene CPS3, phylogenetic analysis and tree, quantitative expression analysis, functional expression of MvCPS3 alone or with MvELS (multifunctional betaalpha-domain class I diterpene synthase) or MvEKS (ent-kaurene synthase) in engineered Nicotiana benthamiana |
| 5.5.1.12 | gene SmCPS1, phylogenetic analysis, a 289-bp gene-specific sequence including the 3' UTR of SmCPS1 is amplified by PCR using cDNA as a template and then cloned into the pK7GWIWG (II) binary, then introduced into Agrobacterium tumefaciens strain EHA105 by electroporation for ransformation of leaf explants from Salvia miltiorrhiza bh2-7 plants, recombinant expression of His-tagged enzyme in Escherichia coli strains Tuner (DE3) or Origami B (DE3) |
| 5.5.1.12 | gene SmCPS2, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strains Tuner (DE3) or Origami B (DE3) |
| 5.5.1.12 | gene TPS-LAS, recombinant expression in Saccharomyces cerevisiae strain AM94, a yeast strain engineered for the production of isoprenoids, which contains three chromosomally integrated copies of a degradation stabilized variant of HMG2 and a heterozygous deletion of ERG9, and in strain AM204, each with or without coexpression of yeast protein Erg20p(F96C), a yeast FPP synthase, overview |
| 5.5.1.12 | Site-directed mutagenesis is carried out via PCR amplification with overlapping mutagenic primers, and the mutant genes verify by complete sequencing. The resulting wild type and mutant genes are then transferred via directional recombination to the T7-promoter and N-terminal 6xhis fusion expression vector pDEST17. Use of the pDEST17 vector results in a 25 amino acid residue linker between the 6 His-tag and the cloned protein. |
