Environmental Genomics Division, National Environmental Engineering Research Institute, CSIR, Nehru Marg, Nagpur, 440020 Maharashtra India.
Indian J Microbiol. 2012 Dec;52(4):538-43. doi: 10.1007/s12088-012-0263-1. Epub 2012 Mar 25.
Metagenomic libraries herald the era of magnifying the microbial world, tapping into the vast metabolic potential of uncultivated microbes, and enhancing the rate of discovery of novel genes and pathways. In this paper, we describe a method that facilitates the extraction of metagenomic DNA from activated sludge of an industrial wastewater treatment plant and its use in mining the metagenome via library construction. The efficiency of this method was demonstrated by the large representation of the bacterial genome in the constructed metagenomic libraries and by the functional clones obtained. The BAC library represented 95.6 times the bacterial genome, while, the pUC library represented 41.7 times the bacterial genome. Twelve clones in the BAC library demonstrated lipolytic activity, while four clones demonstrated dioxygenase activity. Four clones in pUC library tested positive for cellulase activity. This method, using FTA cards, not only can be used for library construction, but can also store the metagenome at room temperature.
宏基因组文库开创了放大微生物世界的新纪元,挖掘了未培养微生物的巨大代谢潜力,并提高了新基因和途径的发现速度。本文描述了一种从工业废水处理厂的活性污泥中提取宏基因组 DNA 的方法,并通过文库构建来挖掘宏基因组。通过构建的宏基因组文库中细菌基因组的大量代表性和获得的功能克隆,证明了该方法的效率。BAC 文库代表了细菌基因组的 95.6 倍,而 pUC 文库代表了细菌基因组的 41.7 倍。BAC 文库中有 12 个克隆显示出脂肪酶活性,而 4 个克隆显示出双加氧酶活性。pUC 文库中的 4 个克隆对纤维素酶活性呈阳性。这种使用 FTA 卡的方法不仅可用于文库构建,还可在室温下存储宏基因组。