引用本文:	李友训,李富超,秦　松,曾志刚,秦蕴珊.东太平洋深海沉积物中DNA的提取及细菌多样性初步分析[J].海洋科学,2008,32(12):69-74.

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东太平洋深海沉积物中DNA的提取及细菌多样性初步分析
李友训^1,2, 李富超¹, 秦　松¹, 曾志刚¹, 秦蕴珊¹
1.中国科学院海洋研究所;2.中国科学院研究生院

摘要:

以东太平洋海隆附近深海柱状沉积物为材料,通过化学裂解和酶消化相结合的方法提取了沉积物微生物的总基因组DNA ,并进行了纯化。结果表明所得到的DNA分子片段大小在21 kb左右,纯化后的DNA可直接用于PCR等分子生物学操作。细菌16S rDNA V3可变区的PCR-DGGE 图谱展示出15条以上条带,表明深海沉积物中细菌多样性较高,群落结构比较复杂。对其中9条主要条带进行回收、测序和系统发育分析,结果表明所获得的序列分属放线菌门(Actinobacteria), 绿弯菌门(Chloroflexi) ,γ-变形细菌亚门( Gam-ma-proteobacteria) ,α-变形细菌亚门(Alpha-proteobacteria) 和嗜酸菌门(Acidobacteria) 5个大类群。

关键词: 深海沉积物 DNA DGGE

DOI：

分类号:

基金项目:中国科学院知识创新工程项目(2007-1 , KZCX3-SW-223,KSCX-2-YW-G-022) ;中国大洋协会项目(DYXM-115-02-2-17)

DNA extraction and DGGE analysis of the bacterial community of a deep-sea sediment sample collected at the East Pacific Ocean

Abstract:

The total genomic DNA of a deep-sea sediment sample collected at the East Pacific Ocean was ext racted using a combined chemical and protease-lysis method. The size of DNA was about 21 kb, and could be used directly for PCR-based molecular research after purification. PCR-DGGE profile of bacterial 16S rDNA V3 fragments gave rise to more than 15 bands. Blast analysis revealed that most of the retrieved sequences from DGGE bands were closely related to uncultured clones from various marine sediment environments, and the dominant groups were Actinobacteria, Chloroflexi, Gamma-proteobacteria , Alpha-proteobacteria and Acidobacteria.

Key words: deep-sea sediment DNA DGGE