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Tetrastigma hemsleyanum ThF3'5' H gene and application thereof

A cloverleaf, gene technology, applied in the direction of application, genetic engineering, plant genetic improvement, etc., to achieve the effect of improving antibacterial properties

Pending Publication Date: 2021-12-31
HANGZHOU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no gene research report related to this active ingredient

Method used

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  • Tetrastigma hemsleyanum ThF3'5' H gene and application thereof
  • Tetrastigma hemsleyanum ThF3'5' H gene and application thereof
  • Tetrastigma hemsleyanum ThF3'5' H gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Acquisition of genes

[0023] 1. Metabolome analysis to obtain differential genes

[0024] Clover leaf samples were frozen, dried, and pulverized into powder for metabolite isolation.

[0025] Take 100mg sample powder and leaching overnight at 4°C with 1.0ml 70% methanol aqueous solution. The extract was pipetted (CNWBOND Carbon GCB SPE Cartridge, 250 mg, 3 ml; Anpel Shanghai, China, www.ANPEL.com.cn / cnw) and filtered (SCAA-104, 0.22 μm pore size; Anpel Shanghai, China, http: / / www .anpel.com.cn / ), followed by LC-MS analysis.

[0026] Sample extracts were analyzed using LC-ESI-MS / MS system (HPLC, gasket set UFLC Shimadzu CBM30A system, www.SHIMADZU.com.cn; MS, Applied Biosystems 6500QTRAP, www.appliedbiosystems.com.cn). Waters ACQUITY UPLC HSS T3 C18 (1.8 μm, 2.1 mm*100 mm) was used for compound separation.

[0027] The analysis conditions are as follows: solvent system, water (0.04% acetic acid): acetonitrile (0.04% acetic acid); gradient program, 95:5 V / V at 0 m...

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Abstract

The invention discloses a tetrastigma hemsleyanum ThF3'5' H gene and application thereof. The nucleotide sequence of the gene is as shown in SEQ ID NO. 1. The application comprises the following steps of firstly, detecting differential expression of tetrastigma hemsleyanum gene in different functional states by utilizing a transcriptome sequencing technology, and determining candidate genes; and cloning the full length of the gene through 3'-RACE and 5'-RACE technologies, finding gene function classification through homologous comparison in a common database of the gene and an amino acid sequence, and finally transforming arabidopsis thaliana through the gene, and detecting the change of metabolite of the transgenic arabidopsis thaliana to determine the function of the gene. It is found that the ThF3 '5' H gene of tetrastigma hemsleyanum can increase the content of flavonoid metabolites kaempferol-3-O-rutin and isorhamnetin-3-O-beta-D-rutinoside of the arabidopsis thaliana plant, and a basis is provided for subsequent research on improvement of the antibacterial property of the arabidopsis thaliana plant.

Description

technical field [0001] The invention relates to the technical field of functional genomics, in particular to a trifoliate ThF3'5'H gene and its application. Background technique [0002] Flavonoids are a class of plant secondary metabolites initiated by phenylpropanoids, which have been extensively studied. Structurally speaking, flavonoids include 2-phenylchromans (2-phenylchromans) and 3-phenylbenzopyrans (3-phenylchromans). 2-Phenylbenzopyrans are flavonoids, including flavanones, flavones, flavonols, flavanols, and anthocyanidins;3 - Phenylbenzopyrans, ie isoflavonoids, including isoflavones, isoflavans and pterocarpans. [0003] The related genes in the synthetic metabolic pathway of flavonoids can be divided into two categories: one is the structural gene encoding the enzyme, and the other is the regulatory gene. Flavanone 3-hydroxylase (flavanone 3-β-hydroxyalse, F3H) catalyzes the C-3 hydroxylation of (2S)-flavanone or (2S)-5-deoxyflavanone synthesized by CHI to g...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/84C12N1/21A01H5/00A01H6/20
CPCC12N9/0073C12N15/8243C12N15/8251C12N15/8279C12N15/8205C12Y114/13088
Inventor 黄雨晴严建立阮松林钱丽华应武陆秋君裘劼人王贤波
Owner HANGZHOU ACAD OF AGRI SCI
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