Microbial technologies for the discovery of novel bioactive metabolites
Microbial technologies for the discovery of novel bioactive metabolites
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DOI：
10.1016/s0168-1656(02)00209-2
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发表时间：
2002-11-13
影响因子：
4.1
通讯作者：
Puglia, AM
中科院分区：
文献类型：
--
作者：
Donadio, S;Monciardini, P;Puglia, AM
土壤微生物是生物活性化合物的重要来源。这些分子具有独特且意想不到的结构，是其分子靶点的选择性抑制剂。在意大利生物搜索公司（Biosearch Italia），新的生物活性分子的发现主要是通过利用一个拥有超过50000株菌株的专有菌株库来进行的，其中大部分是不常见的放线菌属和罕见的丝状真菌。在基于微生物提取物的药物研发中，一个关键因素是分离未被开发的微生物群体，这些微生物同时是次生代谢产物的良好生产者。分子遗传学可以在这些工作中提供帮助。我们将综述分子方法的发展和应用，这些方法用于检测土壤DNA中不常见的放线菌属，以及用于放线菌分离株的快速去重复。结果表明，在许多土壤中不常见的菌属大量存在，并且分离出的放线菌具有高度多样性。然而，尽管不常见的放线菌菌株可能增加产生新结构的机会，但人们对它们的遗传学和生理学了解甚少。为了加快对它们的操作，我们已经开发出了一些载体，这些载体能够在大肠杆菌中稳定地维持放线菌DNA的大片段，并能在链霉菌基因组中进行位点特异性整合。这些载体适用于从克隆DNA的较小片段重建基因簇，从不常见的放线菌菌株制备大插入片段文库以及构建环境文库。（C）2002年由爱思唯尔科学出版社（Elsevier Science B.V.）出版
Soil microbes represent an important source of biologically active compounds. These molecules present original and unexpected structure and are selective inhibitors of their molecular targets. At Biosearch Italia, discovery of new bioactive molecules is mostly carried out through the exploitation of a proprietary strain collection of over 50000 strains, mostly unusual genera of actinomycetes and uncommon filamentous fungi. A critical element in a drug discovery based on microbial extracts is the isolation of unexploited groups of microorganisms that are at the same time good producers of secondary metabolites. Molecular genetics can assist in these efforts. We will review the development and application of molecular methods for the detection of uncommon genera of actinomycetes in soil DNA and for the rapid dereplication of actinomycete isolates. The results indicate a substantial presence in many soils of the uncommon genera and a large diversity of isolated actinomycetes. However, while uncommon actinomycete strains may provide an increased chance of yielding novel structures, their genetics and physiology are poorly understood. To speed up their manipulation, we have developed vectors capable of stably maintaining large segments of actinomycete DNA in Escherichia coli and of integrating site specifically in the Streptomyces genome. These vectors are suitable for the reconstruction of gene clusters from smaller segment of cloned DNA, the preparation of large-insert libraries from unusual actinomycete strains and the construction of environmental libraries. (C) 2002 Published by Elsevier Science B.V.