中文摘要：

为进一步研究紫杉二烯合成酶的作用机理和紫杉醇生物合成代谢调控机制,采用RT-PCR技术从东北红豆杉（Taxus cuspidata）树叶中获得了紫杉二烯合成酶基因片段,将该片段克隆在载体pGEMT-easy vector上,并转化到E.coliJM109中,经EcoRI酶切鉴定后,对阳性克隆进行序列测定,获得紫杉二烯合成酶基因片段长度为909 bp,编码303个氨基酸,与GenBank中收录的紫杉二烯合成酶的同源性达到98%;对获得的紫杉二烯合成酶基因和紫杉醇产生菌HQD33基因组DNA进行了Southern杂交。结果初步证实了紫杉二烯合成酶存在于通过内生真菌发酵生产紫杉醇的生物合成途径中。为利用分子生物学手段研究通过内生真菌发酵生产紫杉醇生物合成的代谢调控和构建高产紫杉醇基因工程菌株提供了强有力的理论指导。

英文摘要：

In order to verify whether taxadiene synthase also exists in the metabolic pathway of taxol production by microbe fermentation or not,southern hybridization between taxol-producing fungus HQD33DNA and taxadiene synthase cDNA from cultured Taxus cuspidata cells were designed.The results showed that the cloned taxadiene synthase gene was 909 bp,encoding 303 amino acids and has 98% identity with reported taxadiene synthase gene sequences and HQD33 genomic DNA could hybridize with the taxadiene synthase probe.This demonstrates that the taxadiene synthase exists in the taxol biosynthesis pathway of endofungus fermentation.These results provide the basis for studies on metabolizing regulation of taxol production by endofungus fermentation and construction of engineering strains with high taxol output by molecular biology methods.