武夷菌素部分生物合成基因簇的克隆和分析  

Cloning and Analysis of Wuyiencin Partial Biosynthetic Gene Cluster of Streptomyces ahygroscopicus var. wuyiensis CK-15

作　　者：葛蓓孛[1] 杨振娟[1] 檀贝贝[1] 刘彦彦[1] 刘艳[1] 孙蕾[1] 张克诚[1] 

机构地区：[1]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

出　　处：《中国生物防治学报》2014年第5期678-684,共7页Chinese Journal of Biological Control

基　　金：公益性行业(农业)科研专项(201103002;201303025);国家自然科学基金(31371985;31301710)

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。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 synthetase gene, which was used for screening CK-15 genomic library. Nine positive clones were identified from the Streptomyces ahygroscopicus var. wuyiensis CK-15 fosmid genomic library. The positive clones were sequenced. There were three large scaffolds with approximately 53.291 kb of gene sequence. This sequence contains 14 possible ORFs and show high homology with nystatin biosynthetic gene of S. noursei ATCC 11455. The research will facilitate the further functional analysis of the wuyiencin biosynthetic genes and breed high yield new strain through genetic engineering.

关 键 词：武夷菌素 生物合成基因簇 克隆 分析 

分 类 号：S476[农业科学—农业昆虫与害虫防治]