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Applied and Environmental Microbiology
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Microbial Ecology

Cloning the Soil Metagenome: a Strategy for Accessing the Genetic and Functional Diversity of Uncultured Microorganisms

Michelle R. Rondon, Paul R. August, Alan D. Bettermann, Sean F. Brady, Trudy H. Grossman, Mark R. Liles, Kara A. Loiacono, Berkley A. Lynch, Ian A. MacNeil, Charles Minor, Choi Lai Tiong, Michael Gilman, Marcia S. Osburne, Jon Clardy, Jo Handelsman, Robert M. Goodman
Michelle R. Rondon
Department of Plant Pathology, University of Wisconsin-Madison, Madison, Wisconsin;
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Paul R. August
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Alan D. Bettermann
Department of Plant Pathology, University of Wisconsin-Madison, Madison, Wisconsin;
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Sean F. Brady
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York
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Trudy H. Grossman
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Mark R. Liles
Department of Plant Pathology, University of Wisconsin-Madison, Madison, Wisconsin;
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Kara A. Loiacono
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Berkley A. Lynch
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Ian A. MacNeil
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Charles Minor
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Choi Lai Tiong
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Michael Gilman
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Marcia S. Osburne
ARIAD Pharmaceuticals, Inc., Cambridge, Massachusetts; and
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Jon Clardy
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York
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Jo Handelsman
Department of Plant Pathology, University of Wisconsin-Madison, Madison, Wisconsin;
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Robert M. Goodman
Department of Plant Pathology, University of Wisconsin-Madison, Madison, Wisconsin;
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DOI: 10.1128/AEM.66.6.2541-2547.2000
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ABSTRACT

Recent progress in molecular microbial ecology has revealed that traditional culturing methods fail to represent the scope of microbial diversity in nature, since only a small proportion of viable microorganisms in a sample are recovered by culturing techniques. To develop methods to investigate the full extent of microbial diversity, we used a bacterial artificial chromosome (BAC) vector to construct libraries of genomic DNA isolated directly from soil (termed metagenomic libraries). To date, we have constructed two such libraries, which contain more than 1 Gbp of DNA. Phylogenetic analysis of 16S rRNA gene sequences recovered from one of the libraries indicates that the BAC libraries contain DNA from a wide diversity of microbial phyla, including sequences from diverse taxa such as the low-G+C, gram-positive Acidobacterium,Cytophagales, and Proteobacteria. Initial screening of the libraries in Escherichia coli identified several clones that express heterologous genes from the inserts, confirming that the BAC vector can be used to maintain, express, and analyze environmental DNA. The phenotypes expressed by these clones include antibacterial, lipase, amylase, nuclease, and hemolytic activities. Metagenomic libraries are a powerful tool for exploring soil microbial diversity, providing access to the genetic information of uncultured soil microorganisms. Such libraries will be the basis of new initiatives to conduct genomic studies that link phylogenetic and functional information about the microbiota of environments dominated by microorganisms that are refractory to cultivation.

  • Copyright © 2000 American Society for Microbiology
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Cloning the Soil Metagenome: a Strategy for Accessing the Genetic and Functional Diversity of Uncultured Microorganisms
Michelle R. Rondon, Paul R. August, Alan D. Bettermann, Sean F. Brady, Trudy H. Grossman, Mark R. Liles, Kara A. Loiacono, Berkley A. Lynch, Ian A. MacNeil, Charles Minor, Choi Lai Tiong, Michael Gilman, Marcia S. Osburne, Jon Clardy, Jo Handelsman, Robert M. Goodman
Applied and Environmental Microbiology Jun 2000, 66 (6) 2541-2547; DOI: 10.1128/AEM.66.6.2541-2547.2000

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Cloning the Soil Metagenome: a Strategy for Accessing the Genetic and Functional Diversity of Uncultured Microorganisms
Michelle R. Rondon, Paul R. August, Alan D. Bettermann, Sean F. Brady, Trudy H. Grossman, Mark R. Liles, Kara A. Loiacono, Berkley A. Lynch, Ian A. MacNeil, Charles Minor, Choi Lai Tiong, Michael Gilman, Marcia S. Osburne, Jon Clardy, Jo Handelsman, Robert M. Goodman
Applied and Environmental Microbiology Jun 2000, 66 (6) 2541-2547; DOI: 10.1128/AEM.66.6.2541-2547.2000
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KEYWORDS

bacteria
Ecosystem
Genome, Bacterial
soil microbiology

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