https://scholars.lib.ntu.edu.tw/handle/123456789/446102
標題: | A novel retrieval system for nearly complete microbial genomic fragments from soil samples | 作者: | Yu W.-H. Su S.-C. CHIA-YIN LEE |
關鍵字: | Metagenomic library; Microbial community structure | 公開日期: | 2008 | 卷: | 72 | 期: | 2 | 起(迄)頁: | 197-205 | 來源出版物: | Journal of Microbiological Methods | 摘要: | The construction of a complex genomic library is one of the comprehensive ways to study a complex bacterial community and to access the variety of metabolic pathways present in the rich soil environment. In this report, we developed a new protocol whereby we are able to retrieve nearly complete microbe genomic fragments from soil samples, which are employed to generate a metagenomic library for visualizing the basic scaffolding of the soil microbial community. The use of direct cell lysis within soil-embedded agarose plugs, along with a double-size selection, enabled us to successfully isolate pure and high-molecular weight DNA (0.1-1 Mb) without the need for any further purification. A metagenomic library containing 1.2 Gbp of DNA in total was constructed. Furthermore, analysis of the microbial community structure using 16S rDNA partial sequences found the dominant phylotypes to consist of α-Proteobacteria and Actinobacteria, which are similar to those seen in forest and agricultural soils, and numerous uncultured microbes from a wide variety of bacterial taxa as well. In conclusion, this study presents a novel protocol for generating a metagenomic library that carries much larger and diverse DNA fragments from soil bacteria that will be applied for the reconstruction of soil microbial genomes and the discovery of novel habitat-specific pathways. © 2007 Elsevier B.V. All rights reserved. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/446102 | ISSN: | 01677012 | DOI: | 10.1016/j.mimet.2007.11.022 | SDG/關鍵字: | agarose; DNA 16S; DNA fragment; Actinobacteria; agricultural land; Alphaproteobacteria; article; DNA determination; DNA library; DNA purification; DNA sequence; forest soil; genome analysis; habitat; microbial community; microbial genetics; molecular phylogeny; molecular weight; nonhuman; nucleotide sequence; priority journal; soil microflora; taxonomy; Bacteria; Chromosomes, Artificial, Bacterial; DNA, Bacterial; Genes, rRNA; Genetic Techniques; Genome, Bacterial; Genomic Library; Molecular Sequence Data; Phylogeny; RNA, Ribosomal, 16S; Soil Microbiology; Actinobacteria; Alphaproteobacteria; Bacteria (microorganisms) |
顯示於: | 農業化學系 |
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