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Taxol is a tetracyclic diterpenoid amides Wani et al 1971 extracted from Taxus brevifolia (Taxus brevifolia) bark, roots and branches, is a highly efficient, low toxicity, broad-spectrum natural anti-cancer drugs, paclitaxel natural extracts derived from the skin of the yew plant (Taxus), and its content is only 0.01% to 0.05%. Taxus are endangered species, growth is very slow, traditional methods need to cut down a large number of yew trees, whichever bark extract paclitaxel, its costly It is estimated that for every 300 trees, about 10 tons of bark can be extracted about 1kg of paclitaxel, about serving five patients; therefore, to seek new ways of paclitaxel, is the protection of the yew and the key to reduce the cost of drug treatment with paclitaxel this production-related areas, such as chemical synthesis, cell culture plant tissue and microbial fermentation and so did a lot of research work. Microbial fermentation, separation and identification of paclitaxel endophytic fungi show people a new way of production paclitaxel. Isolated from different tissues of the wild Chinese yew (Taxus chinensis (Pilger) Rehd.) To 123 endophytic fungi found 4 production Taxanes endophytic fungi detected by HPLC, were named Cbr31 -1, Cbr34-1, Cbr37-2 and Cbr51-2. Using the fungi of ITS rDNA of universal primers object sequence, isolates CBR1-1 Cbr31-1, Cbr31-3, Cbr34-1, Cbr37-2, Cbr38-2, Cbr44-3, Cbr46-2 and Cbr51-2 and the ITS sequences of PCR amplification, and PCR products were sequenced these nine isolates were preliminary species classified Phylogenetic analysis. Optimized chromatographic conditions, the use of high performance liquid natural growth of Taxus chinensis, the Chinese yew Taxus cuspidata shoots 3 Taxanes content detection and analysis. Three kinds of taxane-type compound: 10 - de-acetylated baccatin III (10-deacetyl Baccatin III), baccatin III (baccatin III) and paclitaxel (Taxol) is placed under the same chromatographic conditions detected, to compare different content types taxanes, laid the basis for the development and utilization of resources for the Taxus endophytic fungi producing Taxanes fermentation products to establish a platform for extraction and detection. Fermentation conditions to produce taxol genetically engineered strains EFY-21, and experiment with different basal medium, temperature, pH and other factors on genetically engineered bacteria EFY-21 comparison experiments to determine the optimum culture conditions, the growth curve Determination and optimization of culture conditions, to determine the best medium of carbon and nitrogen sources, and orthogonal experiments the optimum proportion of carbon and nitrogen, and laid the foundation for the future use of genetically modified endogenous fungal large-scale production of paclitaxel.
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