海洋放线菌NRPS基因的筛选  

NRPS gene screening of marine Actinomycetes

作　　者：陈路劼[1] 赵薇[1] 林如[1] 谢阳[1] 江红[1] 连云阳[1] 

机构地区：[1]福建省微生物研究所,福建省新药(微生物)筛选重点实验室,福州350007

出　　处：《海峡药学》2012年第9期240-243,共4页Strait Pharmaceutical Journal

基　　金：福建省自然科学基金项目〔2010〕01081国家重大专项"重大新药创新"药物筛选及相关技术平台(2009ZX9302-004)

摘　　要：目的建立快速、高效的NRPS基因筛选体系,从海洋微生物中筛选具有潜在合成多肽类次级代谢产物的活性菌株,为定向获得新的含氨基酸抗生素奠定基础。方法采用选择性培养分离获得海洋放线菌,通过快速抗菌活性初筛,设计特异性引物利用PCR技术对初筛活性菌株进行NRPS基因筛选,进一步对阳性菌株进行16S rDNA排重。结果利用建立的NRPS基因筛选体系对初筛具有抗菌活性的295株放线菌进行筛选,得到12株阳性菌株,分别属于小单孢菌属、链霉菌属和疣孢菌属。结论利用PCR技术建立了快速、高效的NRPS基因筛选体系,为多肽类活性化合物的获得提供了基因学的依据。OBJECTIVE Establish a rapid,highly efficient screening system for NRPS gene to gain strains potentially producing peptide containing antibiotics.Laying the groundwork for identifying novel peptide-type antibiotics.METHODS Rapid antimicrobial screening was performed for positive strains from marine Actinomycetes isolated,then screened for NRPS gene by PCR amplification using specific primers.16S rDNA sequences of positive strains were analysed for classification and to exclude the replicas.RESULTS 295 strains,out of 3150 marine Actinomycetes isolated,showed antimicrobial activity.Among them,12 bioactive strains were obtained with positive NRPS gene and belonged to Micromonospora,Streptomyces and Verrucosispora,respectively.CONCLUSION A rapid,effective NRPS gene screening system was established using PCR technique.It provides genetics envidence for acquiring novel peptide antibiotics.

关 键 词：海洋放线菌 NRPS 基因筛选 

分 类 号：R914[医药卫生—药物化学]