The Identification Of Taxol-related Enzyme Genes In Taxus Chinensis Endophyte A. Niger

Taxol is a new kind of anti-caneer drug,It has been a focus of the research and development to produce and exploit the taxol by various methods.The good advantage of microbiological fermentation to produce taxol is much more superior than other methods.This test is to produce taxol as the research object which from the northeast yew fungus aspergillus Niger A.n iger.,this paper discusses the synthesis of taxol related enzyme inducer,precursor(that is,the synthesis of taxol way related enzyme substrate)inhibitor and metabolic pathways of A.n iger synthesis of taxol content regulation function;This experiment obtained GGPPS gene fragment from endophytic fungus A.niger with RT-PCR technique,providing a theoretical basis for the synthesis of metabolic regulation mechanism and metabolic pathways of microbial fermentation to produce taxol The main results of the experiment were listed below:1,The syntheses of taxol was significantly promoted by adding some inductors,100.0mg/Lmethyl-jasmonate,20mg/Lsalicylic-acid,1.5mg/Lsilver-nitrate,100mg/L chitosan oligosaccharide,respectively;adding precusor substance together with10.0mg/L PHE,20.0mg/L Ser,0.6mg/L Na Ac and 15mg/L sodium benzoate;adding some Metabolic pathways inhibitors : 2mg/L Gibberellic Acid,15mg/LCinnamic acid,respectively;into the S-7 medium with an appropriate concentration;2,The gene fragment of ggpps was cloned from endophytic fungus A.niger.The sequence was analyzed by Blast programon,obtained fragment size is 371 bp,encoding 108 aa.Gene Bank comparing result showed that the sequence was cloned in the experiment had highly homologous with sequenees(98%)collected from Gene Bank;3,It was verified that ggpps also existed in the biosynthesis pathway of using endophytic fungus A.niger synthesis anti-drug taxol with fermentation method.