Tripterygium wilfordii cytochrome P450s catalyze the methyl shift and epoxidations in the biosynthesis of triptonide - Nature Communications

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Tripterygium wilfordii cytochrome P450s catalyze the methyl shift and epoxidations in the biosynthesis of triptonide - Nature Communications
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. Parent strains without functional URA3 were made competent by the following procedure: Inoculation from a glycerol stock into 5 ml YPD medium and growing at 30 °C O/N. Then, transfer of 3 mL of O/N culture to 50 mL YPD medium and continued growing for 4–5 h followed by centrifugation at 3500 ×for 10 min, then discarding the supernatant. Cells were then ready for transformation after 2 washes in sterile water and resuspension in 0.4 mL of sterile water.

. For colony PCR, colonies were resuspended in 50 µL 20 mM NaOH and incubated at 99 °C for 15 min. 1 µL colony suspension was used for PCR.strain was transferred into 0.5 mL media in a 96-well plate and grown for 3 d at 30 °C with orbital shaking at 350 rpm. For extraction, 0.1 mL ofS. cerevisiae

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