利用KS基因筛选Ⅱ型聚酮类化合物产生菌的方法研究  

The method for screening type Ⅱ polyketide-producting strains based on the KS gene

作　　者：姜华[1] 李晶[1,2] 闫建芳[2] 胡英畅[2] 齐小辉[2] 刘秋[2] 

机构地区：[1]辽宁师范大学生命科学学院,辽宁大连116029 [2]大连民族学院生命科学学院,辽宁大连116600

出　　处：《辽宁师范大学学报（自然科学版）》2010年第1期96-101,共6页Journal of Liaoning Normal University:Natural Science Edition

基　　金：国家自然科学基金项目(30671398);NSFC-KOSEF(韩国科学与工程基金会)项目(30711140389);辽宁省自然科学基金项目(20062189);辽宁省教育厅科学技术研究项目(2008S135;2004F079);大连市青年基金项目(2005J22JH040)

摘　　要：根据酮基合酶编码基因（ketoacyl synthase，KS）的同源性设计简并引物，并利用PCR技术快速筛选携带聚酮类化合物编码基因的放线菌菌株．试验筛选了33株放线菌菌株，检测获得PKSⅡ型阳性菌株16株．克隆并测序获得16条KS基因序列，进行BLAST同源性比对，与GenBank中已知的KS基因序列的相似度在89％～99％之间．提交GenBank，获得的各菌株KS基因登录号为FJ620885～FJ620889，FJ620892，FJ878801～FJ878810．抑菌实验表明，携带KS基因的16株放线菌中，13株具有抑制真菌活性，2株具有抑制革兰氏阳性细菌活性．系统进化树分析表明16株放线菌可以分为7个类群，其中，具有产生新獭聚酮类化合物潜力的菌株8株，说明筛选出的KS基因具有一定的多样性．结果表明，利用PCR技术可以快速有效筛选Ⅱ型KS基因，为生物资源的开发利用莫定基础．An efficient and rapidly method for strains screening was evaluated in this paper. New degenerate PCR primers were designed based on the homology of type Ⅱ KS gene. PCR was used to screen the strains which include type Ⅱ ketoacyl synthase（KS）. In this test,33 actinomycetes strains were screened,and 16 of them were verified that they carried type Ⅱ KS genes. Cloned and sequenced, then the 16 KS sequences were obtained,do BLAST research and the BLAST results indicated that the similarity between these sequences and GenBank was 89%-99%. We submitted the 16 sequences to GenBank and got their accession numbers FJ620885-FJ620889,FJ620892,FJ878801-FJ878810. The bacteriostatic experiment showed that 13 strains had antifungi activity and 2 strains had resistent gram-positive bacteria. Phylogenetic tree showed that the 16 strains could be divided into 7 groups and 8 strains have a potential in producting new polyketide,which may prove that the selected KS genes have a certain degree of diversity. The study indicated that PCR could be used to detect type Ⅱ KS gene rapidly and effectively, thus it laid the foundation for the development and utilization of biological resources all over the world.

关 键 词：放线菌 聚酮类化合物 酮基合酶 抑菌活性 

分 类 号：Q785[生物学—分子生物学] Q939.13