Construction of a metagenomic library for the marine sponge Halichondria okadai

Abe T., Sahin F. P., Akiyama K., Naito T., Kishigami M., Miyamoto K., ...More

Bioscience, Biotechnology and Biochemistry, vol.76, no.4, pp.633-639, 2012 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 76 Issue: 4
  • Publication Date: 2012
  • Doi Number: 10.1271/bbb.110533
  • Journal Name: Bioscience, Biotechnology and Biochemistry
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.633-639
  • Keywords: 454-pyrosequencing, Fosmid, Halichondria okadai, Metagenomic library, Sponge
  • Anadolu University Affiliated: No


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.