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Applied and Environmental Microbiology, March 2008, p. 1649-1652, Vol. 74, No. 5
0099-2240/08/$08.00+0     doi:10.1128/AEM.01864-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Rapid DNA Library Construction for Functional Genomic and Metagenomic Screening{triangledown} ,{dagger}

Jonathan E. Schmitz,1,2 Anu Daniel,1 Mattias Collin,1,{ddagger} Raymond Schuch,1 and Vincent A. Fischetti1*

The Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, 1230 York Avenue, Box 172, New York, New York 10021,1 Weill Cornell/Rockefeller/Sloan-Kettering Tri-Institutional M.D.-Ph.D. Program, 1300 York Avenue, Suite C-103, New York, New York 100212

Received 3 August 2007/ Accepted 1 December 2007

A rapid protocol was developed for constructing plasmid libraries from small quantities of genomic/metagenomic DNA. The technique utilizes linker amplification with topoisomerase cloning and allows for inducible transcription in Escherichia coli. As proof of principle, several anti-Bacillus lysins were cloned from bacteriophage genomes and an aerolysin was cloned from a metagenomic sample.

* Corresponding author. Mailing address: Laboratory of Bacterial Pathogenesis and Immunology, Rockefeller University, 1230 York Avenue, Box 172, New York, NY 10021. Phone: (212) 327-8166. Fax: (212) 327-7584. E-mail: vaf{at}rockefeller.edu

{triangledown} Published ahead of print on 14 December 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

{ddagger} Present address: Department of Clinical Sciences, Division of Infection Medicine, Lund University, Klinikgatan 26, SE-221 84 Lund, Sweden.

Applied and Environmental Microbiology, March 2008, p. 1649-1652, Vol. 74, No. 5
0099-2240/08/$08.00+0     doi:10.1128/AEM.01864-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Waschkowitz, T., Rockstroh, S., Daniel, R. (2009). Isolation and Characterization of Metalloproteases with a Novel Domain Structure by Construction and Screening of Metagenomic Libraries. Appl. Environ. Microbiol. 75: 2506-2516 [Abstract] [Full Text]  


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