Mutation Breeding Of Endophytic Fungi Of Taxol And Analysis Of Its Genetic Difference

It is believed that the taxol is a kind of anti-cancer drug, and it is very effective for many kinds of cancer. Taxol was initially extracted from the taxus brevifolia and with microtubule depolymerization stabilizer novel mechanism of action. As the bark of taxus paclitaxel extracted far from being able to meet the needs of the market, And taxus belonging to the endangered plants,therefore we must find other ways.It is showed that using genetic engineering on the basis of microbial fermentation can construct high-yielding Taxol strain.This experiment to Taxol-producing strain HD86-9 as the starting strain,use the method of UV mutagenesis and Licl composite UV mutagenesis to make the bacteria mutagenesis, through the improved S-7 fermentation medium, thin-layer chromatography for the production of detection qualitative, high-performance liquid chromatography for the output of quantitative measurement, and ultimately selected high-yield strain of taxol. The mutation rate and lethallty rate show that optimum mutagenesis time of Taxol-producing strain HD86-9 in 6% Licl is 7.5～8.5 min behind the 15W ultraviolet lamp 30cm.Screen 5 high-yield strain of taxol based on HPLC result. The yield of Taxol is 274.76μg/L,and 35.6% more than the HD86-9.HD86-9 and high-yield strains produced Taxol of the cDNA as a template amplify using 4 anchor primer and 10 arbitray primer. SDS-Page detected the different expressed gene fragments, sequencingd and analyzing the gene fragments.This study established the foundation of isolating HD86-9 taxol biosynthesis ralated genes,clarifying taxol metabolic pathway, Laid the groundwork of construcing high-yield strain of taxol.