Construction of soil environmental DNA cosmid libraries and screening for clones that produce biologically active small molecules
Construction of soil environmental DNA cosmid libraries and screening for clones that produce biologically active small molecules
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DOI：
10.1038/nprot.2007.195
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Brady, Sean F.
中科院分区：
文献类型：
--
作者：
Brady, Sean F.
对环境样品的非培养研究表明，大多数细菌不容易在实验室中培养。从环境样品中成功培养出的一小部分细菌是一种非常有价值的新型生物活性天然产物的来源。将直接从环境样品中提取的DNA引入容易培养的细菌中，并筛选这些大型文库中产生生物活性小分子的克隆，是获得由先前未培养细菌的基因组编码的天然产物的一种手段。该协议提供了详细的程序克隆DNA直接从环境样品和筛选这些克隆的抗菌活性的天然产物的生产。从土壤样本到鉴定具有抗菌活性的环境DNA克隆，整个过程大约需要两周时间。
Culture-independent studies on environmental samples indicate that most bacteria are not readily cultured in the laboratory. The small fraction of bacteria that have been successfully cultured from environmental samples have been a very rewarding source of novel biologically active natural products. The introduction of DNA extracted directly from environmental samples into easily cultured bacteria and the screening of these large libraries for clones that produce biologically active small molecules is one means to access natural products encoded by the genomes of previously uncultured bacteria. This protocol provides detailed procedures for cloning DNA directly from environmental samples and screening these clones for the production of antibacterially active natural products. The entire protocol, from soil sample to the identification of antibacterially active environmental DNA clones, will take approximately 2 weeks.