Construction of a Metagenomic Library for the Marine Sponge <I>Halichondria okadai</I>

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  • ABE Takahiro
    Department of Chemistry, Faculty of Science, Kanagawa University Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University Department of Chemistry, Faculty of Science, Kanagawa University Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University
  • SAHIN Fatma Pinar
    Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University Department of Pharmaceutical Botany, Faculty of Pharmacy, Hacettepe University
  • AKIYAMA Kiyotaka
    Molecular Neuroscience Unit, Okinawa Institute of Science and Technology Molecular Neuroscience Unit, Okinawa Institute of Science and Technology
  • NAITO Takayuki
    Molecular Neuroscience Unit, Okinawa Institute of Science and Technology Molecular Neuroscience Unit, Okinawa Institute of Science and Technology
  • KISHIGAMI Mizoe
    Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University
  • MIYAMOTO Kenji
    Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University
  • SAKAKIBARA Yasufumi
    Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University
  • UEMURA Daisuke
    Department of Chemistry, Faculty of Science, Kanagawa University Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University Department of Chemistry, Faculty of Science, Kanagawa University Department of Biosciences and Informatics, Faculty of Science and Technology, Keio University

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  • Construction of a Metagenomic Library for the Marine Sponge Halichondria okadai
  • Construction of a metagenomic library for the marine dponge Halichondria okadai

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Abstract

Symbionts of the marine sponge Halichondria okadai are promising as a source of natural products. Metagenomic technology is a powerful tool for accessing the genetic and biochemical potential of bacteria. Hence, we established a method of recovering bacterial-enriched metagenomic DNA by stepwise centrifugation. The metagenomic DNA was analyzed by ultrafast 454-pyrosequencing technology, and the results suggested that more than three types of bacterial DNA, Alphaproteobacteria, Actinobacteria, and Cyanobacteria, had been recovered, and that eukaryotic genes comprised only 0.02% of the metagenomic DNA. These results indicate that stepwise centrifugation and real-time quantitative PCR were effective for separating sponge cells and symbiotic bacteria, and that we constructed a bacteria-enriched metagenomic library from a marine sponge, H. okadai, selectively for the first time.

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