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High coverage sequencing of DNA from microorganisms living in an oil reservoir 2.5 kilometres subsurface
Author(s) -
Kotlar Hans K.,
Lewin Anna,
Johansen Jostein,
ThroneHolst Mimmi,
Haverkamp Thomas,
Markussen Sidsel,
Winnberg Asgeir,
Ringrose Philip,
Aakvik Trine,
Ryeng Einar,
Jakobsen Kjetill,
Drabløs Finn,
Valla Svein
Publication year - 2011
Publication title -
environmental microbiology reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.229
H-Index - 69
ISSN - 1758-2229
DOI - 10.1111/j.1758-2229.2011.00279.x
Subject(s) - metagenomics , pyrosequencing , microorganism , biology , escherichia coli , enolase , dna , 16s ribosomal rna , microbial enhanced oil recovery , microbial population biology , gene , bacteria , genetics , immunohistochemistry , immunology
Summary Microorganisms colonize a variety of extreme environments, and based on cultivation studies and analyses of PCR‐amplified 16S rDNA sequences, microbial life appears to extend deep into the earth crust. However, none of these studies involved comprehensive characterizations of total DNA. Here we report results of a high‐coverage DNA pyrosequencing of an apparently representative and uncontaminated sample from a deep sea oil reservoir located 2.5 km subsurface, attributing a pressure and temperature of 250 bars and 85°C respectively. Bioinformatic analyses of the DNA sequences indicate that the reservoir harbours a rich microbial community dominated by a smaller number of taxa. Comparison of the metagenome with sequences in databases indicated that there may have been contact between the oil reservoir and surface communities late in the sequence of geological events leading to oil reservoir formation. One specific gene, encoding a putative enolase, was synthesized and expressed in Escherichia coli . Enolase activity was confirmed and was found to be much more thermotolerant than for a corresponding E. coli enzyme, consistent with the conditions in the oil reservoir.