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PEDS Advance Access originally published online on May 31, 2007
Protein Engineering Design and Selection 2007 20(6):267-271; doi:10.1093/protein/gzm019
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© The Author 2007. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Using archaeal histones for precise DNA fragmentation

E. Azzoni1, D. Sblattero2, M. Licciulli1, R. Marzari1 and P. Edomi1,3

1 Department of Biology, University of Trieste, via Giorgieri 7, I-34127 Trieste, Italy 2 Department of Medical Sciences, University of Eastern Piedmont, via Solaroli 17, 28100 Novara, Italy

3 To whom correspondence should be addressed. E-mail: edomi{at}units.it

The fragmentation of DNA is a useful procedure for many molecular biology procedures. However, most methods used to fragment DNA are poorly controllable, and cannot be used to create small fragments. We describe a method to generate random DNA fragments of a predictable size to be cloned in expression vectors for the construction of display libraries. The DNA is allowed to form complexes with archaeal histones from Methanothermus fervidus (HMf) and the HMf/DNA core complex is naturally protected from nuclease DNaseI activity, giving rise to DNA fragments of ~60 bp and multiples thereof. We found that by varying the wt/wt ratio between DNA and HMf, the concentration of DNA and the incubation time with DNaseI, DNA fragments of desired size can be obtained. This approach should be applicable to the efficient fragmentation of DNA for the construction of phage display polypeptide libraries, as well as any other molecular biology procedures in which small DNA fragments of defined size are required.

Keywords: archaeal histones/DNA fragmentation/expression gene libraries/phage display

Received December 21, 2006; revised March 28, 2007; accepted April 12, 2007.


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