Method for effectively inhibiting taxus media cell biosynthesis C-14 oxygenated taxane

A technology of Taxus mandya and taxane, which is applied in biochemical equipment and methods, determination/inspection of microorganisms, introduction of foreign genetic material using vectors, etc., can solve problems such as reducing synthesis

Inactive Publication Date: 2010-07-21
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

There are no reports at home and abroad on inhibiting the expression of taxane 14β-hydroxylase gene in Taxus x media cells and reducing the synthesis of C-14 oxidized taxanes in Taxus x media cells

Method used

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  • Method for effectively inhibiting taxus media cell biosynthesis C-14 oxygenated taxane
  • Method for effectively inhibiting taxus media cell biosynthesis C-14 oxygenated taxane
  • Method for effectively inhibiting taxus media cell biosynthesis C-14 oxygenated taxane

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Embodiment 1

[0028] Embodiment 1: Use the method for effectively inhibiting the biosynthesis of C-14 oxidized taxanes in Taxus x media (Taxus x media) cells of the present invention, and target the taxus taxane 14β hydroxylase gene (abbreviated 14OH) The antisense gene expression vector constructed from the fragment was introduced into the cultured cells of Taxus x media. Through the optimized transformation method, the expression of the 14OH gene in the Taxus mandiae cells and the biosynthesis of three -14 oxidized taxanes, sinenxan A, sinenxan C, and Yunnanxane, in the Taxus mandiaea cells were significantly inhibited.

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Abstract

The invention relates to a method for effectively reducing taxus media cell biosynthesis C-14 oxygenated taxane, which belongs to the technical field of genetic engineering. Through an meloidogynosis agrobacterium mediated binary vector transformation system, the method leads an antisense genetic expression vector constructed according to yew taxane 14Beta hydroxylase gene (referred to as 14OH) fragments into a Taxus media cultured cell. The method successfully obtains transgenic cells through an optimized transformation method. PCR-Southern hybridization, RT-PCR gene expression analysis and chemical component detection results have shown that the taxus media cell transgenic test succeeds, and the 14OH gen mRNA expression level in a transgenic cell system is significantly reduced. The method significantly inhibits the flowing of the taxus media cells to C-14 oxygenated taxoids, and is expected to lead metabolism to flow to C-13oxygenated taxoids including an anti-cancer drug taxol and the precursor baccatin III thereof, so as to provide a novel method for improving the biosynthesis yield of taxol.

Description

technical field [0001] The invention relates to a method for effectively reducing the synthesis of C-14 oxidized taxanes by Taxus x media cells. It belongs to the technical field of genetic engineering developed in the 1980s on the basis of plant cell culture. As a new technology, plant cell suspension culture technology can produce useful drugs on a large scale, and the prospect is very promising. Background technique [0002] Paclitaxel (taxol) is an important secondary metabolite isolated from the bark of Taxus Brevifolia for the first time by Wall et al. It can be used clinically to treat ovarian cancer, lung cancer, bladder cancer, uterine cancer and AIDS-related Karposis sarcoma, etc. It is one of the most widely used natural anticancer drugs at present. However, currently paclitaxel is mainly extracted from the yew plant, and the 2000-pound bark of 2000-3000 yew trees is required to produce 1 kg of paclitaxel. Due to over-cutting, the wild resources of yew have bee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/82C12Q1/68C12Q1/02G01N30/02
Inventor 邱德有李凤岚胡新玲
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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