Promoter for regulating expression of genes in T shape trichome and application of promoter
A non-secretory glandular trichome and gene regulation technology, applied in the field of promoters, can solve the problems of temporal and spatial regulation of target genes, excessive consumption of cellular material and energy, and harm
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[0038] This embodiment relates to the acquisition of the AaTTPS5 gene promoter, which specifically includes the following steps:
[0039] Step 1. Cultivate aseptic seedlings of Artemisia annua
[0040] Artemisia annua seeds were soaked in ethanol with a volume fraction of 75% for 1 min, then soaked in 20% (w / v) NaClO for 20 min, rinsed with sterile water for 3 to 4 times, blotted the surface moisture with sterile absorbent paper, and inoculated Cultivate on MS solid medium without any hormone, 25°C, 16h / 8h (light / dark) light culture, after 14 days, you can obtain the sterile vaccine of Artemisia annua;
[0041] Step 2: Cloning of the promoter sequence in the genomic DNA
[0042] 1. Genomic DNA Extraction
[0043] Add 2 steel balls into a 1.5mL centrifuge tube, and put a piece of Artemisia annua leaf (1cm 2 About the size, take it in an ice box). Add 300 μL TPS buffer (operate in a fume hood, TPS contains 2% mercaptoethanol), shake at 55-60 Hz for 90 seconds. Then add 300 ...
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