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Genosenoside biological synthesis relative gene

A technology for biosynthesis and ginsenoside is applied in the field of genetic engineering for improving plant traits, and can solve the problems of complex chemical structure and synthesis process of ginsenoside, complicated and expensive extraction and preparation, and inability to chemically synthesize ginsenoside, and shorten the biosynthesis cycle. Effect

Inactive Publication Date: 2004-09-29
XIANGYA MEDICAL COLLEGE CENT SOUTH UNIV
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Problems solved by technology

However, because the content of each ginsenoside monomer in ginseng is very small, such as Rg3, which has an obvious anti-tumor metastasis effect, is only 3 / 100,000, and the extraction and preparation are extremely complicated and expensive. Saponins still cannot be chemically synthesized
At present, a small amount of ginsenosides is mainly extracted from cultivated ginseng with very low content, but it is difficult to meet the huge demand for clinical medicine
Therefore, the very small content of ginsenosides in plant ginseng has become a bottleneck restricting the development of new drugs for ginsenoside monomers

Method used

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Embodiment Construction

[0008] 1. Isolation of RNA from root tissues of four-year-old and one-year-old ginseng plants and purification of mRNA

[0009] (1) Using the guanidinium isothiocyanate method to isolate high-quality RNA from four-year-old and annual fresh and clean ginseng plant root tissues;

[0010] (2) Take 200 μg of isolated four-year-old and one-year-old ginseng root tissue RNA respectively, and use PolyATractmRNA Isolation System (Promega) to purify mRNA.

[0011] 2. Synthesis of cDNA from root tissues of 4-year-old and 1-year-old ginseng plants

[0012] The first strand of cDNA and the second strand of cDNA were synthesized by reverse transcription using the mRNA of root tissue of four-year-old and one-year-old ginseng plants respectively.

[0013] (1) The reaction system for the first strand synthesis of cDNA is as follows:

[0014] mRNA 5μl (2μg)

[0015] cDNA synthesis primer (10μmol / L) 1μl

[0016] 5×1st strand buffer 2μl

[0017] 10mM dNTPs 1μl

[0018] AMV reverse transcrip...

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Abstract

The present invention relates to gene engineering of improving plant characteristics. By means of inhibiting degressive hybridization and other process, six genesenoside biological synthesis relative genes are cloned, including GPR5 with cDNA sequence length of 421 bp, GBR6 with cDNA sequence legnth of 1049 bp, GBR1 with cDNA sequence legnth of 145 bp, GBR2 with cDNA sequence length of 227 bp, GBR3 with cDNA sequence length of 291 bp and GBR4 with cDNA sequence length of 400 bp. Through constituting efficient expression and conversion vector containing the above said genes and ginseng tissue culture of transgenic plant to produce gensenoside, the present invention lays the foundation for raising the amount of biologically synthesized Rg3, Rb1, Rg1 and gensenoside monomers and shortening their biologically synthesizing period.

Description

technical field [0001] The invention relates to genetic engineering for improving plant traits, in particular to promoting genes related to ginsenoside biosynthesis. Background technique [0002] Among the medicinal plants, ginseng (Panax ginseng C.A. Meyer) belongs to Araliaceae. It is a traditional precious Chinese medicinal material with world influence. It has a history of thousands of years in my country and Southeast Asia. It is a very typical medicinal plant. and representative ginseng plants. Ginsenoside is its main medicinal active ingredient, composed of panaxadiol and panaxatriol-type saponins, belonging to tetracyclic triterpene dammarane saponins, Compounds composed of isoprene units (5 carbons) (see the simplified chemical structure formula), so far, more than 40 species of saponins have been isolated and determined from the plant ginseng. In recent years, through the monomer The pharmacological activity of saponin components has been thoroughly studied, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07H21/04C12N15/29C12P19/34
Inventor 罗志勇陆秋恒胡维新陈湘晖罗建清刘水平
Owner XIANGYA MEDICAL COLLEGE CENT SOUTH UNIV
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