title: Whole-genome sequencing of unculturable bacterium using whole-genome amplification.
abstract: More than 99% of microorganisms on the earth are unculturable with known culturing techniques. The emergence of metagenomics with high-throughput sequencing technologies has enabled researchers to capture a comprehensive view of a complex bacterial community which comprises both culturable and unculturable species. However, the function of an individual species remains difficult to elucidate in a conventional metagenomic study, which generates numerous genomic fragments of unidentifiable origins at a species or genus level. This limitation hampers any in-depth investigations of the community and its unculturable bacterial members. Recently, as an alternative or compensatory approach, genomics targeting a single unculturable bacterial species in a complex community has been proposed. In this approach, whole-genome amplification technique using Phi29 DNA polymerase is applied to obtain a sufficient quantity of DNA for genome sequence analysis from only a single to a thousand bacterial cells. It is expected that a combination of the conventional metagenomics and this single-species-targeting genomics provides a great progress in understanding of the ecology, physiology, and evolution of unculturable microbial communities.
authors: Department of Biological Sciences, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Tokyo, Japan. yhongo@bio.titech.ac.jp
authors: Yuichi
authors: Y
authors: Hongoh
authors: Atsushi
authors: A
authors: Toyoda
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics* / Q000254:growth & development; D046508:Culture Techniques; D016680:Genome, Bacterial / Q000235:genetics*; D023281:Genomics / Q000379:methods*; D020049:Isoptera / Q000382:microbiology; D055616:Microtechnology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*; D013559:Symbiosis