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