A kind of α-santalene synthase, gene and application
A technology of santalene and synthetase, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., and can solve the problem of low product specificity
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Embodiment 1
[0019] Example 1: Preparation of α-santalene synthase, gene, vector and engineered bacteria derived from Turmeric
[0020] 1. Homology analysis of candidate CwSS genes
[0021] The tuberous root, tender leaves, buds and stem tissues were collected from both sides of Feiyun River in Ruian, Wenzhou, and the transcriptome was sequenced by Shanghai Meiji Biotechnology Company. Bioinformatics analysis of the obtained sequences revealed a candidate sesquiterpene synthase gene ( That is, the α-santalene synthase of the present application, CwSS), its deduced amino acid sequence was subjected to Blastp analysis at NCBI, and it was found that it had the greatest similarity with the bisabolene synthase (Accession Number: D2YZP9) derived from Zingiber officinale. source, the sequence identity reaches 86%; further sequence alignment with other sources of sesquiterpene synthase shows that CwSS has conserved domains, such as DDxxD, RXR, NSE / DTE, RRX 8 W( figure 1 ).
[0022] Phylogenetic...
Embodiment 2
[0028] Example 2: Analysis of Catalytic Properties of Recombinase CwSS
[0029] The CwSS recombinase solution (concentration is 50mg / L, volume is 10μL) prepared by the method of Example 1 is used as catalyst, pH 7.0 Tris-HCl buffer solution, 2μg FPP (farnesyl pyrophosphate), 500mM MgCl are added. 2 20 μL of the solution, 2 μL of 500 mM DTT, and 100 μL of dilute glycerol (1,2,3-propanetriol) constitute 1 ml of the reaction system, respectively, and fully react at 30 °C for 120 min. After the reaction was completed, the extraction head was taken out and injected into a gas chromatograph to conduct qualitative analysis of the product. Chromatographic conditions: GC-2010 Shimadzu gas chromatograph; chromatographic column HP-5; carrier gas: N 2 , the purge flow is 3mL / min, no split; the initial temperature of the column oven is 40°C, and it is kept for 2 minutes, and then it is heated to 220°C at a rate of 7°C / min, and it is kept for 5 minutes; the inlet temperature is 250°C; The...
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