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Molecular biologic techniques applied to the microbial prospecting of oil and gas in the Ban 876 gas and oil field in China

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Abstract

Currently, molecular biologic techniques achieve a great development in studies of soil samples. The objective of this research is to improve methods for microbial prospecting of oil and gas by applying culture-independent techniques to soil sampled from above a known oil and gas field. Firstly, the community structure of soil bacteria above the Ban 876 Gas and Oil Field was analyzed based on 16S rRNA gene clone libraries. The soil bacteria communities were consistently different along the depth; however, Chloroflexi and Gemmatimonadetes were predominant and methanotrophs were minor in both bacteria libraries (DGS1 and DGS2). Secondly, the numbers of methane-oxidizing bacteria, quantified using a culture-dependent procedure and culture-independent group-specific real-time PCR (RT-PCR), respectively, were inconsistent with a quantify variance of one or two orders of magnitude. Special emphasis was given to the counting advantages of RT-PCR based on the methanotrophic pmoA gene. Finally, the diversity and distribution of methanotrophic communities in the soil samples were analyzed by constructing clone libraries of functional gene. All 508-bp inserts in clones phylogenetically belonged to the methanotrophic pmoA gene with similarities from 83% to 100%. However, most of the similarities were below 96%. Five clone libraries of methanotrophs clearly showed that the anomalous methanotrophs (Methylosinus and Methylocystis) occupy the studied area.

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Acknowledgments

This study was sponsored by the Petrotech Junior Innovation Fund of China (code: 07E1025). The authors wish to thank Qingxian Feng, a Senior Engineer of the DaGang Oil Field, for kindly providing geo-materials from the studied gas and oil field.

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Authors and Affiliations

  1. The Key Laboratory of Marine Reservoir Evolution and Hydrocarbon Accumulation Mechanism, Ministry of Education, China; School of Energy Resources, China University of Geosciences (Beijing), Beijing, 100083, China

    Fan Zhang & Dujie Hou

  2. College of Chemistry and Environmental Engineering, Yangtze University, Hubei, 434023, China

    Yuehui She & Shuqiong Kong

  3. State Key Laboratory of Urban and Regional Ecology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China

    Yong Zheng & Zhifeng Zhou

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  1. Fan Zhang
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  2. Yuehui She
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  3. Yong Zheng
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  4. Zhifeng Zhou
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  5. Shuqiong Kong
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  6. Dujie Hou
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Correspondence to Yuehui She or Dujie Hou.

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Zhang, F., She, Y., Zheng, Y. et al. Molecular biologic techniques applied to the microbial prospecting of oil and gas in the Ban 876 gas and oil field in China. Appl Microbiol Biotechnol 86, 1183–1194 (2010). https://doi.org/10.1007/s00253-009-2426-5

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  • DOI: https://doi.org/10.1007/s00253-009-2426-5

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