Flavanone 3',5'-hydroxylase gene and protein coded by same
A flavanone and hydroxylase technology, applied in the field of genetic engineering, can solve the problem of not finding such genes, etc., and achieve the effect of improving ornamental quality
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[0013] 1. Cloning of candidate gene fragments in the flower of Aconitum sichuanensis using merger primers
[0014] In Aconitum aconitum (a well-known public material, Qu Suping et al. "Rapid Propagation of Aconitum Aconitum Seedlings by Tissue Culture" Chinese Wild Plant Resources CHINESE WILD PLANT RESOURCES 2004,23(4):62-63) flowering period, whichever is different developmental period After the flowers were quickly frozen in liquid nitrogen, the total RNA was extracted with TRIZOL (invitogen) according to the reagent instructions. Transcribe it into cDNA, then dilute the cDNA 10 times as a template, and carry out PCR reaction with degenerate primers. The names and sequences of the degenerate primers are as follows:
[0015] W-FH-F: 5'-C(A / T / C / G)CA(A / G)GA(T / C)ATGGT(T / C / G)TT(C / T)G-3'
[0016] W-FH-R: 5’-C(A / T / G)AT(A / T / C / G)GCCCA(A / T / G)AT(A / G)TT
[0017] (A / T / C / G)A-3'
[0018] 0.5 μl each of 10 μM 5’ primer and 3’ primer; 2.5 μl 10× buffer; 2.5 μl 2.5 mM dNTP; 1.5 μl 25 mM m...
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