Isolation of High Molecular Weight DNA from Marine Sponge Bacteria for BAC Library Construction
Ouyang, YC; Dai, SK; Xie, LW; Kumar, MSR; Sun, W; Sun, HM; Tang, DL; Li, X; lixiang@scsio.ac.cn
2010
发表期刊MARINE BIOTECHNOLOGY
ISSN1436-2228
卷号12期号:3页码:318-325
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.
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
关键词Dna Recovery Marine Sponge Bacterial Artificial Chromosome Metagenomic Library
学科领域Biotechnology & Applied Microbiology ; Marine & Freshwater Biology
URL查看原文
收录类别sci
资助项目LMB
WOS记录号WOS:000278095400010
引用统计
被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/3824
专题中科院海洋生物资源可持续利用重点实验室
通讯作者lixiang@scsio.ac.cn
推荐引用方式
GB/T 7714
Ouyang, YC,Dai, SK,Xie, LW,et al. Isolation of High Molecular Weight DNA from Marine Sponge Bacteria for BAC Library Construction[J]. MARINE BIOTECHNOLOGY,2010,12(3):318-325.
APA Ouyang, YC.,Dai, SK.,Xie, LW.,Kumar, MSR.,Sun, W.,...&lixiang@scsio.ac.cn.(2010).Isolation of High Molecular Weight DNA from Marine Sponge Bacteria for BAC Library Construction.MARINE BIOTECHNOLOGY,12(3),318-325.
MLA Ouyang, YC,et al."Isolation of High Molecular Weight DNA from Marine Sponge Bacteria for BAC Library Construction".MARINE BIOTECHNOLOGY 12.3(2010):318-325.
条目包含的文件
文件名称/大小 文献类型 版本类型 开放类型 使用许可
Isolation of High Mo(216KB) 开放获取--请求全文
个性服务
推荐该条目
保存到收藏夹
查看访问统计
导出为Endnote文件
相关权益政策
暂无数据
收藏/分享
所有评论 (0)
暂无评论
 

除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。