Evaluation of the Use of Amplified 16S rRNA Gene-restriction Fragment Length Polymorphism Analysis to Detect Enterobacter cloacae and Bacillus licheniformis for Microbial Enhanced Oil Recovery Field Pilot
Evaluation of the Use of Amplified 16S rRNA Gene-restriction Fragment Length Polymorphism Analysis to Detect Enterobacter cloacae and Bacillus licheniformis for Microbial Enhanced Oil Recovery Field Pilot
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DOI：
10.1627/jpi1958.42.342
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发表时间：
1999-09
影响因子：
1
通讯作者：
K. Fujiwara;Tanaka Shinji;Makiko Ohtsuka;N. Ichimura;H. Yonebayashi;C. Hong;H. Enomoto
中科院分区：
文献类型：
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作者：
K. Fujiwara;Tanaka Shinji;Makiko Ohtsuka;N. Ichimura;H. Yonebayashi;C. Hong;H. Enomoto
对注入油藏的微生物的16 S rRNA基因的限制性片段长度多态性（RFLP）分析的有效性进行了评价，以监测它们随时间的水平。本研究从供试微生物中筛选出2种微生物，即阴沟肠杆菌TRC-322和地衣芽孢杆菌TRC-18-2-a，采用聚合酶链反应（PCR）技术，以1对通用引物扩增其16 S rRNA基因片段。采集储层卤水和储层岩石样品，分离出储层中的微生物，在相同条件下扩增其16 S rRNA基因。根据需要，使用限制性内切酶HhaI、MspI、TaqI对这些微生物中的每一种的16 S rRNA基因进行RFLP分析。结果表明，近缘种的RFLP图谱与E. Cloptosis TRC-322或B。地衣型TRC-18-2-a不在从储层分离的微生物中。采用本文提出的方法对16 S rRNA基因进行PCR-RFLP分析，可有效地检测是否存在适当的注射微生物。这种方法对于研究从水库中分离的微生物也是有效的，这些微生物具有在糖蜜上生长的能力。
Evaluation of effectiveness of restriction fragment length polymorphism (RFLP) analysis of the 16S rRNA gene of microorganisms injected into an oil reservoir, for monitoring their levels over time, was conducted. Two microorganisms, Enterobacter cloacae TRC-322 and Bacillus licheniformis TRC-18-2-a, were focused in this paper among the microorganisms selected for injection, and gene fragments of the 16S rRNA gene of these microorganisms were amplified by polymerase chain reaction (PCR), using one set of universal primers. Samples of the reservoir brine and reservoir rock were obtained; the microorganisms inhabiting in the reservoir were isolated from these samples, and the 16S rRNA gene of these microorganisms was amplified, condition remaining the same. RFLP analysis was performed on the 16S rRNA gene of each of these microorganisms, using restriction endonucleases HhaI, MspI, AluI and TaqI as necessary. Comparison of the resultant rRNA gene fragments, demonstrated that closely-related species displaying RFLP profile similar to that of E. cloacae TRC-322 or B. licheniformis TRC-18-2-a were not among the microorganisms isolated from the reservoir. PCR-RFLP analysis of the 16S rRNA gene, using the protocol presented in this paper, is effective to detect the presence of appropriate injecting microorganisms. This method was also effective for studying microorganisms isolated from the reservoir, which have the ability to grow on a molasses.