CAJ | 학술논문

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。
불흡수련매균무이변충Streptomyces ahygroscopicus var. wuyiensis CK-15시종복건성무이산토양중분리득도적일주련매균，기대사산물무이균소대과소진균병해구유량호적방치효과，단시인기산량저적결점한제료무이균소공업화생산화농업생산중적응용。위료실현이용기인공정배육고산신균주적목표，수선요획득무이균소적생물합성기인。근거대배내지류항생소취동합성매기인설계인물사선균주CK-15적기인조문고，공획득9개양성극륭。극륭화측서획득3개교장scaffold편단，서렬총장도체53.291 kb，기중포함료14개가능열독광，통과동원비대증실해서렬여S. noursei ATCC 11455적제매소생물합성기인유흔고적동원성。본연구위진일보연구무이균소생물합성기인적공능，병통과기인공정배육고산신균주전정료기출。
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.