Abstract
The vast majority of the Earth’s biological diversity is hidden in uncultured and yet uncharacterized microbial genomes. The construction of metagenomic libraries is a cultivation-independent molecular approach to assess this unexplored genetic reservoir. In the last few years, a high number of novel biocatalysts have been identified by function-based or sequence-based screening of metagenomic libraries. Here, we describe detailed protocols for the construction of metagenomic small-insert and large-insert libraries in plasmids and fosmids, respectively, from environmental DNA.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Handelsman, J. (2004) Metagenomics: application of genomics to uncultured microorganisms. Microbiol Mol Biol Rev 68, 669–685.
Steele, H.L., Jaeger, K.-E., Daniel, R., and Streit, W.R. (2009) Advances in recovery of novel biocatalysts from metagenomes. J Mol Microbiol Biotechnol 16, 25–37.
Daniel, R. (2005) The metagenomics of soil. Nat Rev Microbiol 3, 470–478.
Riesenfeld, C.S., Schloss, P.D., and Handelsman, J. (2004) Metagenomics: genomic analysis of microbial communities. Annu Rev Genet 38, 525–552.
Daniel, R. (2004) The soil metagenome – a rich resource for the discovery of novel natural products. Curr Opin Biotechnol 15, 199–204.
Jogler, C., Lin, W., Meyerdierks, A., Kube, M., Katzmann, E., Flies, C., Pan, Y., Amann, R., Reinhardt, R., and Schüler, D. (2009) Towards cloning the magnetotactic metagenome: Identification of magnetosome island gene clusters in uncultivated magnetotactic bacteria from different aquatic sediments. Appl Environ Microbiol 75, 3972–3979.
Simon, C., Herath, J., Rockstroh, S., and Daniel, R. (2009) Rapid identification of genes encoding DNA polymerases by function-based screening of metagenomic libraries derived from glacial ice. Appl Environ Microbiol 75, 2964–2968.
Waschkowitz, T., Rockstroh, S., and Daniel, R. (2009) Isolation and characterization of metalloproteases with a noval domain structure by construction and screening of metagenomic libraries. Appl Environ Microbiol 75, 2506–2516.
Zhang, K., Martiny, A.C., Reppas, N.B., Barry, K.W., Malek, J., Chisholm, S.W., and Church, G.M. (2006) Sequencing genomes from single cells by polymerase cloning. Nat Biotechnol 24, 680–686.
GE Healthcare. IllustraTM GenomiPhi V2 DNA Amplification Kit: instruction manual. GE Healthcare, Munich. http://www.gehealthcare.com/usen/index.html
Bioline. SureClean: instruction manual. Bioline, Luckenwalde.http://www.bioline.com/
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press
About this protocol
Cite this protocol
Simon, C., Daniel, R. (2010). Construction of Small-Insert and Large-Insert Metagenomic Libraries. In: Streit, W., Daniel, R. (eds) Metagenomics. Methods in Molecular Biology, vol 668. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-823-2_2
Download citation
DOI: https://doi.org/10.1007/978-1-60761-823-2_2
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-822-5
Online ISBN: 978-1-60761-823-2
eBook Packages: Springer Protocols