Method for producing paclitaxel by using corn plant expression paclitaxel synthase gene system

Abstract

The invention discloses a method for producing paclitaxel by using a system for expressing a paclitaxel synthase gene in a corn plant, belongs to the field of molecular biology and genetic engineering, and particularly relates to a method for producing paclitaxel by using a system for efficiently expressing the paclitaxel synthase gene in the corn plant. The invention aims to obtain a method for producing paclitaxel by a system for efficiently expressing a paclitaxel synthase gene in a corn plant, which comprises the following steps of: cloning genes such as Geranylgeranyl Pyrophorate Synthase, Taxadiene Synthase and Taxadiene 5-alpha Hydroxylase in a paclitaxel synthesis route to a plant binary expression vector pCAMBIA3301 through codon optimization, and performing agrobacterium tumefaciens-mediated transformation on an endosperm callus of the corn in the agricultural area 108, so as to obtain the paclitaxel with the high-efficiency expression of the paclitaxel synthase gene in the corn plant, thereby obtaining the paclitaxel with the high-efficiency expression of the paclitaxel synthase gene in the corn plant. And finally obtaining the transgenic maize regenerated plant for expressing paclitaxel and the precursor substance thereof.

Description

technical field [0001] The invention discloses a method for producing paclitaxel by utilizing a maize plant expressing paclitaxel synthase gene system, belonging to the field of molecular biology and genetic engineering, in particular to a method for producing paclitaxel by maize plant expressing paclitaxel synthase gene system efficiently. Background technique [0002] Paclitaxel (paclitaxel, C47H51NO4,) is a tricyclic diterpenoid secondary metabolite extracted from Taxus genus. Originally obtained from the bark of Pacific yew. Paclitaxel has a unique anti-tumor mechanism. It is an anti-microtubule drug, which can combine with β-tubulin to accelerate the polymerization of proteins, thereby promoting the formation of microtubules and stabilizing the formed microtubules. The cell division cycle stops and apoptosis occurs, and at the same time, it has a radiosensitizing effect, which may cause the cells to stop in the G2 and M phases, and the cells in this period are very sen...

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Problems solved by technology

[0006] It can be seen that the production level of paclitaxel can be significantly increased through the eukaryotic expression system, but maize, which is also a eukaryotic organism, has an upstream metabolic system and various enzyme reaction systems for paclitaxel synthesis, and a complete paclitaxel synthesis has not yet been established. system, corn and yew have similar plant metabolism and synthesis systems, and can be widely planted as field crops. The stems, leaves and fruits on the ground are good raw materials for extracting paclitaxel, and the biomass output of the aboveground parts is large. Completely replace the national rare and protected species such as yew, and solve the problem that paclitaxel cannot be widely extracted due to the limitation of yew raw materials in the market

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