武夷菌素部分生物合成基因簇的克隆和分析.pdfVIP

30(5)678-684 中国生物防治学报 Chinese Journal of Biological Control 2014 年 10 月 武夷菌素部分生物合成基因簇的克隆和分析 葛蓓孛，杨振娟，檀贝贝，刘彦彦，刘 艳，孙 蕾，张克诚* （中国农业科学院植物保护研究所/农作物病虫害生物学国家重点实验室，北京 100193） 摘要：不吸水链霉菌武夷变种Streptomyces ahygroscopicus var. wuyiensis CK-15 是从福建省武夷山土样中 分离得到的一株链霉菌，其代谢产物武夷菌素对果蔬真菌病害具有良好的防治效果，但是因其产量低的缺 点限制了武夷菌素工业化生产和农业生产中的应用。为了实现利用基因工程培育高产新菌株的目标，首先 要获得武夷菌素的生物合成基因。根据大环内酯类抗生素聚酮合成酶基因设计引物筛选菌株CK-15 的基因 组文库，共获得9 个阳性克隆。克隆和测序获得3 个较长scaffold 片段，序列总长度达53.291 kb，其中包含 了14 个可能阅读框，通过同源比对证实该序列与S. noursei ATCC 11455 的制霉素生物合成基因有很高的同 源性。本研究为进一步研究武夷菌素生物合成基因的功能，并通过基因工程培育高产新菌株奠定了基础。 关 键 词：武夷菌素；生物合成基因簇；克隆；分析 中图分类号：S476 文献标识码：A 文章编号：1005-9261 (2014)05-0678-07 Cloning and Analysis of Wuyiencin Partial Biosynthetic Gene Cluster of Streptomyces ahygroscopicus var. wuyiensis CK-15 * GE Beibei, YANG Zhenjuan, TAN Beibei, LIU Yanyan, LIU Yan, SUN Lei, ZHANG Kecheng (State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China) Abstract: A wuyiencin producing strain Streptomyces ahygroscopicus var. wuyiensis CK-15 was isolated and purified from Wuyi mountain soil in Fujian province. Wuyiencin as secondary metabolites has good control effect on fruit and vegetable fungal diseases whereas which is limited on its low production disadvantage in industrial production and agricultural application. In order to achieve the aim of breeding high yield strain by genetic engineering, an attempt to obtain the biosynthetic gene cluster of wuyiencin generated strain was made. In this study, primers were designed according to a sequence of macrolide antibiotic polyketone sy