Method for improving content of artemisinin in sweet wormwood herb through transferring F3H gene
A kind of artemisinin and transgenic technology, which is applied in horticultural methods, genetic engineering, plant gene improvement, etc., can solve the problems of artificial synthesis, such as difficulty, feasibility, and inability to fully meet market demand, and achieve the effect of stabilizing new drug sources
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[0031] Step 1, cloning of Artemisia annua F3H gene
[0032] (1) Extraction of total RNA from Artemisia annua genome
[0033]The total RNA of Artemisia annua genome was extracted using RNA kit from TIANGEN Company. Take 50-100 mg young leaves of Artemisia annua and quickly grind them into powder in liquid nitrogen, add 550 μL of lysis solution RL (mercaptoethanol has been added), and vortex vigorously to mix. Centrifuge at 9800rpm for 5min, transfer 450μL to filter column CS, place CS in a collection tube, centrifuge at 12000rpm for 5 minutes, carefully draw 400μL of the supernatant in the collection tube into an RNase-free centrifuge tube, slowly add 0.5 times the volume of the supernatant mixed with absolute ethanol, transferred to the adsorption column CR3, centrifuged at 12,000 rpm for 1 min, discarded the waste liquid, and put CR3 back into the collection tube. Add 350 μL protein-removing solution RW1 to CR3, centrifuge at 12,000 rpm for 1 min, discard the waste liquid, ...
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