Abe Takahiro, Sahin Fatma Pinar, Akiyama Kiyotaka, Naito Takayuki, Kishigami Mizoe, Miyamoto Kenji, Sakakibara Yasufumi, Uemura Daisuke
Department of Chemistry, Faculty of Science, Kanagawa University, Hiratsuka, Kanagawa, Japan.
Biosci Biotechnol Biochem. 2012;76(4):633-9. doi: 10.1271/bbb.110533. Epub 2012 Apr 7.
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.
海洋海绵冈村氏软海绵的共生菌有望成为天然产物的来源。宏基因组技术是挖掘细菌遗传和生化潜能的有力工具。因此，我们建立了一种通过逐步离心回收富含细菌的宏基因组DNA的方法。利用超快速454焦磷酸测序技术对宏基因组DNA进行分析，结果表明已回收了三种以上类型的细菌DNA，即α-变形菌纲、放线菌和蓝细菌，且真核基因仅占宏基因组DNA的0.02%。这些结果表明，逐步离心和实时定量PCR对分离海绵细胞和共生细菌有效，并且我们首次从海洋海绵冈村氏软海绵中选择性地构建了一个富含细菌的宏基因组文库。
