红树林来源Streptomyces+sp.OUC6819中多烯大环内酯类抗生素的生物合成的研究.pdfVIP

opeF ZH1 ZH2 ZH3 和 双交换同源重组的质粒 、 、 ，通过接合转移方法将重组质 粒ZH3 导入野生株Streptomycessp.OUC6819 之中，获得了opeF基因阻断突变 株。对野生株和opeF基因突变株进行发酵，经HPLC-MS 分析表明：突变株丧 失了多烯大环内酯(分子式C H O )的合成能力，证实了opeF基因是多烯大环 36 58 10 内酯生物合成中的正调控子。 最后，在转录水平上研究了opeF基因对基因簇中其它基因的调控，RT-PCR opeF opeA opeE opeF opeG 结果表明， 基因是以正调控方式调控基因簇中的 、 、 、 、 opeK、opeM和opeN基因，这为进一步进行基因工程改造提供理论支持。 关键词： 关键词： 关关键键词词：：Streptomycessp.OUC6819； 多烯大环内酯类化合物； 基因组文库； PCRtargeting； 同源重组； opeF； 正调控子 Studyonthebiosynthesisofpolyenemacrolidesfrom Studyonthebiosynthesisofpolyenemacrolidesfrom SSttuuddyyoonntthheebbiioossyynntthheessiissooffppoollyyeenneemmaaccrroolliiddeessffrroomm mangrove-derivedStreptomycessp.OUC6819 mangrove-derivedStreptomycessp.OUC6819 mmaannggrroovvee--ddeerriivveeddSSttrreeppttoommyycceesssspp..OOUUC Abstract Abstract AAbbssttrraacctt Streptomyces sp.OUC6819 was isolated from reeds rhizosphere soil in Guangdong province of China, which is able to produce a series of 32 membered polyene macrolides (PEMS) with significant antifungal activities. For the purpose of elucidating the biosynthesis mechanism of polyene macrolides in Streptomyces sp.OUC6819 ,in this thesis, the following experimentswerecarried out: Firstly, growth conditions for Streptomyces sp.OUC6819 were studied and optimized. Streptomyces sp.OUC6819 was inoculated in the different medium (MS, ISP2, ISP4, COM, gause) with salt concentration ranging from 0% to 10%. The results of experimental showed the optimum culture conditions are ISP2, 28 ℃, salt concentration 3.3% and pH=7.2. The antibiotic resistance experiment showed Streptomyces sp.OUC6819 is sensitive to Thiost