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学科主题: Biotechnology & Applied Microbiology; Marine & Freshwater Biology
题名: Isolation of High Molecular Weight DNA from Marine Sponge Bacteria for BAC Library Construction
作者: Ouyang, YC ; Dai, SK ; Xie, LW ; Kumar, MSR ; Sun, W ; Sun, HM ; Tang, DL ; Li, X
通讯作者: lixiang@scsio.ac.cn
关键词: DNA recovery ; Marine sponge ; Bacterial artificial chromosome ; Metagenomic library
刊名: MARINE BIOTECHNOLOGY
发表日期: 2010
卷: 12, 期:3, 页:318-325
收录类别: sci
部门归属: [Li, Xiang] Chinese Acad Sci, Key Lab Marine Bioresources Sustainable Utilizat, Guangdong Key Lab Marine Mat Med LMMM GD, S China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China; [Ouyang, Yongchang; Dai, Shikun; Xie, Lianwu; Kumar, M. S. Ravi; Sun, Wei; Sun, Huimin; Li, Xiang] Chinese Acad Sci, LMB, LMMM GD, S China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China; [Ouyang, Yongchang; Dai, Shikun; Xie, Lianwu; Kumar, M. S. Ravi; Sun, Wei; Sun, Huimin] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China; [Kumar, M. S. Ravi; Tang, Danling] Chinese Acad Sci, LED, RSMEE, S China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China
项目归属: LMB
摘要: Metagenomics is a powerful tool for mining the genetic repositories from environmental microorganisms. Bacteria associated with marine sponges (phylum Porifera) are rich sources of biologically active natural products. However, to date, few compounds are discovered from the sponge metagenomic libraries, and the main reason might be the difficulties in recovery of high molecular weight (HMW) DNA from sponge symbionts to construct large insert libraries. Here, we describe a method to recover HMW bacterial DNA from diverse sponges with high quality for bacterial artificial chromosome (BAC) library construction. Microorganisms concentrated from sponges by differential centrifugation were embedded in agarose plugs to lyse out the HMW DNA for recovery. DNA fragments over 436 kb size were recovered from three different types of sponges, Halichondria sp., Haliclona sp., and Xestospongia sp. To evaluate the recovered DNA quality, the diversity of bacterial DNA comprised in the HMW DNA derived from sponge Halichondria sp. was analyzed, and this HMW DNA sample was also cloned into a shuttle BAC vector between Escherichia coli and Streptomyces sp. The results showed that more than five types of bacterial DNA, i.e., Proteobacteria, Nitrospirae, Cyanobacteria, Planctomycetes, and unidentified bacteria, had been recovered by this method, and an average 100 kb size insert DNA in a constructed BAC library demonstrated that the recovered HMW DNA is suitable for metagenomic library construction.
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WOS记录号: WOS:000278095400010
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内容类型: 期刊论文
URI标识: http://ir.scsio.ac.cn/handle/344004/3824
Appears in Collections:中科院海洋生物资源可持续利用重点实验室_期刊论文

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Ouyang, YC; Dai, SK; Xie, LW; Kumar, MSR; Sun, W; Sun, HM; Tang, DL; Li, X.Isolation of High Molecular Weight DNA from Marine Sponge Bacteria for BAC Library Construction,MARINE BIOTECHNOLOGY,2010,12(3):318-325
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