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Flavone glycosyltransferase protein and coding gene and application thereof

A kind of technology of glycosyltransferase, flavonoid glycoside

Active Publication Date: 2019-08-30
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on Andrographis paniculata flavone glycosyltransferase at home and abroad

Method used

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  • Flavone glycosyltransferase protein and coding gene and application thereof
  • Flavone glycosyltransferase protein and coding gene and application thereof
  • Flavone glycosyltransferase protein and coding gene and application thereof

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Experimental program
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Effect test

Embodiment 1

[0033] Discovery of a flavone glycosyltransferase protein ApUGT3 and its coding gene from Andrographis paniculata

[0034] Transcriptome sequencing was performed using MEJA (methyl jasmonate) induction, and nine candidate glycosyltransferases were found in the leaf transcriptome, all of which were induced by methyl jasmonate. Further prokaryotic expression and functional verification of nine genes revealed that the protein ApUGT3 is a glycosyltransferase that can exclusively participate in the biosynthesis of flavonoid glycosides, but cannot catalyze the biosynthesis of diterpenoids in Andrographis paniculata.

[0035] The amino acid of the glycosyltransferase protein ApUGT3 is shown in sequence 1 of the sequence listing, and its corresponding coding region in cDNA is shown in sequence 2 of the sequence listing.

Embodiment 2

[0037] Functional Verification

[0038] 1. Construction of recombinant plasmids

[0039] 1. Synthesize the double-stranded DNA molecule shown in sequence 2 of the sequence listing, namely the ApUFGT3 gene.

[0040] 2. Using the DNA molecule obtained in step 1 as a template, a primer pair composed of F1 and R1 is used for PCR amplification, and the PCR amplification product is recovered.

[0041] F3: 5'-TCCAGGGGCCCGAATTCGGAATGGATCCCAATGTCGAAG-3';

[0042] R3: 5'-AGTGCGGCCGCAAGCTTGTTACTTTGCTTCATTTTTCTC-3'.

[0043]3. Take the vector HIS-MBP-pET28a, perform double digestion with restriction endonucleases BamHI and SalI, and recover the linearized vector backbone. (Recorded in the following documents: Probing the Single Key Amino Acid Responsible for the Novel Catalytic Function of ent-Kaurene Oxidase Supported by NADPH-CytochromeP450 Reductases in Tripterygium Wilfordii; Frontiers in PlantScience, www.frontiersin.org; October 2017, Volume 8, Article 6).

[0044] 4. Take the P...

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Abstract

The invention relates to the technical field of biology, in particular to flavone glycosyltransferase protein and a coding gene and application thereof. The protein is selected from (a1) or (a2): (a1), protein consisting of amino acid sequences as shown in a sequence 1; and (a2), protein obtained through substitution and / or deletion and / or addition on one or several amino acid residues, having functions of glycosyltransferase and derived from the glycosyltransferase in (a1). The protein can perform glycosylation on flavonoid compounds having different structures through sequencing, identification and separation of transcriptomes, and can perform glycosylation on different hydroxyls of the flavonoid compounds.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a flavone glycosyltransferase protein and its coding gene and application. Background technique [0002] Flavonoid glycosides are a class of natural organic compounds that have a series of important physiological activities and widely exist in nature. Flavonoid glycosides mostly exist in plants in the form of glycosides. For example, one of the secondary metabolites in Andrographis paniculata is flavonoids and their glycosides. In recent years, people pay more and more attention to the antiplatelet and anti-proliferation pharmacological effects of flavonoid glycosides in Andrographis paniculata, which provides opportunities for further development and clinical application of flavonoid glycosides in Andrographis paniculata. Glycosylation is a key modification step that occurs in various biological processes, especially in secondary metabolic pathways, where it alters the stability, ...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/54C12N15/70C12N1/21C12P19/60C12R1/19
CPCC12N9/1051C12N15/70C12P19/60C12Y204/01091
Inventor 黄璐琦高伟李媛
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI