Isolation of High Molecular Weight DNA from Marine Sponge Bacteria for BAC Library Construction
文献类型：期刊论文
| 作者 | Li, X; Tang, DL; Sun, HM; Sun, W; Kumar, MSR; Xie, LW; Dai, SK; Ouyang, YC; lixiang@scsio.ac.cn |
| 刊名 | MARINE BIOTECHNOLOGY
 |
| 出版日期 | 2010 |
| 卷号 | 12期号:3页码:318-325 |
| 关键词 | Dna Recovery Marine Sponge Bacterial Artificial Chromosome Metagenomic Library |
| ISSN号 | 1436-2228 |
| 英文摘要 | 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. |
| 学科主题 | Biotechnology & Applied Microbiology ; Marine & Freshwater Biology |
| URL标识 | 查看原文 |
| 资助项目 | LMB |
| WOS记录号 | WOS:000278095400010 |
| 公开日期 | 2011-07-02 |
| 源URL | [http://ir.scsio.ac.cn/handle/344004/3824]  |
| 专题 | 南海海洋研究所_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 | Li, X,Tang, DL,Sun, HM,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 | Li, X.,Tang, DL.,Sun, HM.,Sun, W.,Kumar, MSR.,...&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 | Li, X,et al."Isolation of High Molecular Weight DNA from Marine Sponge Bacteria for BAC Library Construction".MARINE BIOTECHNOLOGY 12.3(2010):318-325. |
入库方式： OAI收割
来源：南海海洋研究所
浏览0
下载0
收藏0
其他版本
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。