东太平洋深海沉积物中DNA的提取及细菌多样性初步分析  

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

作　　者：李友训[1,2] 李富超[1] 秦松[1] 曾志刚[1] 秦蕴珊[1] 

机构地区：[1]中国科学院海洋研究所,山东青岛266071 [2]中国科学院研究生院,北京100049

出　　处：《海洋科学》2008年第12期69-74,共6页Marine Sciences

基　　金：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)

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21kb左右,纯化后的DNA可直接用于PCR等分子生物学操作。细菌16SrDNAV3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(Gamma-proteobacteria),α-变形细菌亚门(Alpha-proteobacteria)和嗜酸菌门(Acidobacteria)5个大类群。The total genomic DNA of a deep-sea sediment sample collected at the East Pacific Ocean was extracted 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.

关 键 词：深海沉积物 DNA DGGE 

分 类 号：Q93[生物学—微生物学]