Carbon glycosyl transferase DhCGT2 gene in pachyrhizus erosus plant and application

A carbon sugar-based, transferase technology, applied in the direction of plant genetic improvement, transferase, application, etc.

Active Publication Date: 2021-06-15
YUNNAN AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the key enzymes CGTs in the biosynthetic path

Method used

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  • Carbon glycosyl transferase DhCGT2 gene in pachyrhizus erosus plant and application
  • Carbon glycosyl transferase DhCGT2 gene in pachyrhizus erosus plant and application
  • Carbon glycosyl transferase DhCGT2 gene in pachyrhizus erosus plant and application

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Embodiment 1

[0036]Based on the basic functional annotation information of the transcriptome Unigene of Tseudoglycans, CGT candidate genes were screened in the sequencing annotation results. Using the gene of Ggcgt as a clue, through sequence-local BLAST analysis, and then sorting and analyzing the screening results, 11 CGT-type carboglycosyltransferases (TcCGT) were finally found. After a series of work such as cDNA preparation, candidate gene amplification and recovery, homologous recombination, protein expression, in vitro enzyme activity reaction, and HPLC and LC / MS detection, it was finally identified that it can catalyze dihydroxynaringenin C The target candidate carbon glycosyltransferase DhCGT2 gene ( figure 1 ). The steps in each stage of the synthesis of schafutasides and isoschafutasides are as follows:

[0037] (1) Preparation of cDNA template

[0038] The fresh leaf samples of Fructus Glycyrrhiza were taken and frozen in liquid nitrogen for RNA extraction. Total RNA was ex...

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Abstract

The invention relates to a carbon glycosyl transferase DhCGT2 gene in a desmodium heterocarpum plant and application thereof, and belongs to the technical field of biology. The nucleotide sequence of the carbon glycosyl transferase DhCGT2 gene in the desmodium heterocarpum plant is as shown in SEQ ID NO.1, and the total length of the sequence is 1422 bp; and the amino acid sequence of the encoded protein is shown as SEQ ID NO.2, 473 amino acid residues are encoded, and the encoded protein can be used for preparing schaftoside and isoschaftoside. The carbon glycosyl transferase is found in desmodium plant desmodium heterocarpum for the first time, and has the functions of catalyzing the generation of vitexin and isovitexin by taking dyhydroxy naringenin as a substrate and catalyzing the generation of schaftoside and isoschaftoside by taking dyhydroxy naringenin glucose C-glycoside as a substrate; and an efficient biosynthesis method for heterologous expression of vitexin, isovitexin, schaftoside and isoschaftoside in vitro of plants is provided.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a carbon syltransferase DhC GT2 gene in a pseudoglyph plant and its application in the preparation of vitexin, isovitexin, schaftoside and isoschaftoside. Background technique [0002] The medicinal plant Desmodium heterocarpon (L.) DC. belongs to the Leguminosae genus Desmodium. It is distributed both at home and abroad, and it is mainly distributed in Yunnan and Guangdong in China. The whole herb of the false ground bean is used as medicine, which has the effects of relieving pain, clearing heat, diuresis, anti-inflammatory, calming the liver, detoxifying, promoting blood circulation, relieving cough and relieving asthma. According to "Chinese Materia Medica", it is recorded that False Ground Bean has a significant effect on the treatment of epidemic encephalitis B, hepatitis, and mumps. False ground beans mainly contain flavonoid glycosides, alkaloids, terpenes and ot...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/10C12N15/70C12N1/21C12P17/16C12R1/19
CPCC12N9/1048C12N15/70C12P17/162Y02A50/30
Inventor 郝冰袁慧娟张广辉杨生超刘祥宇谢家伟陈小巧
Owner YUNNAN AGRICULTURAL UNIVERSITY
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