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多杀菌素是对农业虫害防治及粮食仓储安全均具有重大意义的农用抗生素。为了深入揭示刺糖多孢菌合成多杀菌素的调控特点,首先通过建立基于报告基因的启动子探测技术,探测了多杀菌素生物合成基因簇的9个启动子活性。并进一步通过荧光定量PCR,分析了这9个基因和不在基因簇内的负责糖基前体供应和鼠李糖合成的4个基因的转录时序,结果表明多杀菌素生物合成基因簇内的9个基因在菌体生长进入稳定期时有较高的转录,这和发酵液中此时开始大量积累多杀菌素一致;同时还发现,簇外的4个与糖基供应相关的基因和基因簇内基因的转录时序不同,它们在菌体生长对数期有较高的转录活性,这暗示多杀菌素聚酮链的合成速率和参与后修饰的糖基底物供应的最优化匹配有可能是提高生物合成多杀菌素的前提和关键。
Spinosad is an agricultural antibiotic that is of great importance to the prevention and control of agricultural pests and the safety of grain storage. In order to further reveal the regulatory features of spinosad synthesis spinosad, we first detected the promoter activity of spinosad biosynthetic gene cluster by establishing reporter-based promoter detection technology. Furthermore, the transcriptional timing of the four genes responsible for the supply of glycosyl precursors and the synthesis of rhamnose in the nine genes and non-gene clusters was analyzed by fluorescence quantitative PCR. The results showed that the spinosad biosynthesis gene cluster 9 The results showed that a large number of genes were transcribed in the stationary phase of the cell growth, which was consistent with the accumulation of spinosad at this time in the fermentation broth. At the same time, four gene and gene cluster The internal genes differ in transcriptional timing and have a high transcriptional activity in the logarithmic phase of the cell growth, suggesting that the optimal rate of spinosad synthesis and the post-modification glycolate supply may be increased Biosynthesis of spinosad premise and key.