石油学报 ›› 2005, Vol. 26 ›› Issue (6): 82-85,89.DOI: 10.7623/syxb200506018

• 油田开发 • 上一篇    下一篇

变性梯度凝胶电泳方法在内源微生物驱油研究中的应用

程海鹰1,2, 肖生科2, 汪卫东2, 张翰奭1, 王修林1   

  1. 1. 中国海洋大学化学化工学院, 山东, 青岛, 266003;
    2. 中国石化胜利油田有限责任公司采油工艺研究院, 山东, 东营, 257000
  • 收稿日期:2004-11-29 修回日期:2005-01-18 出版日期:2005-11-25 发布日期:2010-05-21
  • 作者简介:程海鹰,男,1975年9月生,2003年获西安石油大学硕士学位,现为中国海洋大学化学化工学院在读博士研究生,主要从事微生物采油方面的研究.E-mail:chenghaiying25@163.com
  • 基金资助:
    国家“十五”重大科技攻关项目(2001BA605A-11)部分成果

Application of denaturing gradient gel electrophoresis method to study of oil recovery using indigenous microorganism

CHENG Hai-ying1,2, XIAO Sheng-ke2, WANG Wei-dong2, ZHANG Han-shi1, WANG Xiu-lin1   

  1. 1. College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266003, China;
    2. Oil Production Technology Research Institute, Shengli Oilfield Co., Ltd., Dongying 257000, China
  • Received:2004-11-29 Revised:2005-01-18 Online:2005-11-25 Published:2010-05-21

摘要: 应用基于聚合酶链式反应(PCR)的变性梯度凝胶电泳方法(DGGE)分析了在高温、高压和厌氧条件下富集培养的油藏内源微生物16S rDNA。用化学裂解法(SDS)直接提取了油藏微生物基因组DNA,并以此基因组DNA为模板,用特异性引物对16SrDNA的V8和V9可变区进行聚合酶链式反应扩增。对长约400bp(碱基对)的扩增产物进行DGGE分离后,得到了分离较好的电泳图谱。结果表明,DGGE方法可以分离不同油藏微生物16S rDNA的V8和V9可变区的DNA扩增片断。在内源微生物驱油研究中,DGGE方法是在分子水平上分析油藏微生物群落结构和监测微生物群落动态变化的有效工具。

关键词: 变性梯度凝胶电泳方法, 油藏, 内源微生物, 16SrDNA, 微生物驱油, 微生物基因

Abstract: Denaturing gradient gel electrophoresis method (DGGE) based on polymerase chain reaction (PCR) was used to analyze the indigenous microorganism 16S rDNA that was enriched under high-temperature high-pressure and anaerobic conditions of oil reservoir. Genomic DNA was extracted directly from the enriched samples by SDS. Taking genomic DNA as templates, 16S rDNA fragments in V8 and V9 regions were amplified by PCR with specific ribosomal primers. The fragments of 16S rDNA with 400-bp in length were amplified and then separated by DGGE. Well separated and high-quality profiles presumably representing the dominant bacterial fractions in enriched cultures were obtained. The results showed that DGGE could clearly separate the amplified fragments of 16S rDNA in V8 and V9 regions with different indigenous microorganisms, which makes it possible to further identify and define these 16S rDNA fragments by gene sequencing or other means. In the studies of oil recovery using indigenous microorganisms, DGGE is valuable for analyzing microbial community structures and monitoring community dynamics at the molecular level.

Key words: denaturing gradient gel electrophoresis method, reservoir, indigenous microorganisms, 16S rDNA, microbial enhanced oil recovery, microbial gene

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