title: Construction of a metagenomic library for the marine sponge Halichondria okadai.
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.
authors.affiliation: Department of Chemistry, Faculty of Science, Kanagawa University, Hiratsuka, Kanagawa, Japan. abtaka@kanagawa-u.ac.jp
authors.forename: Takahiro
authors.initials: T
authors.lastname: Abe
authors.forename: Fatma Pinar
authors.initials: FP
authors.lastname: Sahin
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Akiyama
authors.forename: Takayuki
authors.initials: T
authors.lastname: Naito
authors.forename: Mizoe
authors.initials: M
authors.lastname: Kishigami
authors.forename: Kenji
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Sakakibara
authors.forename: Daisuke
authors.initials: D
authors.lastname: Uemura
mesh_terms: D039903:Actinobacteria / Q000235:genetics*; D020561:Alphaproteobacteria / Q000235:genetics*; D000818:Animals; D002498:Centrifugation; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D015698:Genomic Library; D059014:High-Throughput Nucleotide Sequencing; D056186:Metagenomics; D010802:Phylogeny; D011161:Porifera / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013559:Symbiosis / Q000502:physiology
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012337:RNA, Ribosomal, 18S