The gene encoding the anthocyanin reductase of S. chinensis and its application
A technology of wisteria anthocyanin and reductase, which is applied to the gene encoding spaghetti anthocyanin reductase and its application field, can solve the problems of undiscovered research and reports, and achieve the effect of high application value
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Embodiment 1
[0031] (1) RNA and DNA extraction: Trizol reagent (Invitrogen, USA) was used to extract total RNA from fresh young leaves of Mildew Spatholobus, and pretreated with RNase Free DNase (Promega, USA) to eliminate genomic DNA contamination. RNA integrity was analyzed on a 1.5% agarose gel, and RNA purity and concentration were determined spectrophotometrically.
[0032] The total DNA was extracted from the young leaves of Mildew Spatholobus by plant DNA kit (TIANGEN).
[0033] (2) Gene cloning: design primers according to the ANR sequence in the Genome of Spatholobus Spatholobus, use DNA as a template, and use Primer5.0 software to design gene-specific primers
[0034] F1: CGGATTTATGGCCTCTTG (SEQ ID NO: 4)
[0035] R1: TGACTTGATTCTCGCACC (SEQ ID NO: 5)
[0036] Through PCR amplification, a gDNA sequence with a length of 1996 bp was obtained, such as SEQ ID NO:1.
Embodiment 2
[0039] The RNA extracted in Example 1 was reverse-transcribed (AMV first strand cDNA synthesis kit: Roche, Switzerland), using the first strand cDNA as a template, using primers:
[0040] F1:TCTTTCCAGCTTGCTACACC (SEQ ID NO: 6)
[0041] R1: CTTTGAGGGACAATCTTCG (SEQ ID NO: 7)
[0042] Through PCR amplification, the cDNA sequence SEQ ID NO: 2 with a length of 644bp was obtained.
Embodiment 3
[0044] According to the translation of the cDNA sequence in Example 2, the amino acid sequence SEQ ID NO: 3 encoding anthocyanidin reductase (SsANR) of S. santiculata was obtained.
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