Promoter for regulating expression of genes in T shape trichome and application of promoter
A non-secretory glandular hair, gene regulation technology, applied in the field of promoters, can solve the problems of time and space regulation of target genes, excessive consumption of cellular substances and energy, harm, etc.
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[0038] This embodiment relates to the acquisition of the AaPDS1 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 a hormone-free MS solid medium at 25°C under 16h / 8h (light / dark) light, and after 14 days, you can obtain an Artemisia annua sterile vaccine;
[0041] Step 2: Cloning of the promoter sequence in the genomic DNA
[0042] 1. Genomic DNA extraction
[0043] Put a leaf of Artemisia annua (1cm 2 About the size, take it in an ice box), add 2 steel balls. Add 300 μL TPS buffer (operate in a fume hood, TPS contains mercaptoethanol), shake at 55-60 Hz for 90 seconds. Then add 300 μL TPSbuffer (fume hood). Water bath at 65°C for...
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