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Ultrasound-mediated DNA transfer for bacteria

Song, Yizhi; Hahn, Thomas; Thompson, Ian P.; Mason, Timothy J.; Preston, Gail M.; Li, Guanghe; Paniwnyk, Larysa; Huang, Wei E. 2007 Ultrasound-mediated DNA transfer for bacteria. Nucleic Acids Research, 35 (19), e129. 10.1093/nar/gkm710

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Abstract/Summary

In environmental microbiology, the most commonly used methods of bacterial DNA transfer are conjugation and electroporation. However, conjugation requires physical contact and cell–pilus–cell interactions; electroporation requires low-ionic strength medium and high voltage. These limitations have hampered broad applications of bacterial DNA delivery. We have employed a standard low frequency 40 kHz ultrasound bath to successfully transfer plasmid pBBR1MCS2 into Pseudomonas putida UWC1, Escherichia coli DH5 and Pseudomonas fluorescens SBW25 with high efficiency. Under optimal conditions: ultrasound exposure time of 10 s, 50 mM CaCl2, temperature of 22°C, plasmid concentration of 0.8 ng/µl, P. putida UWC1 cell concentration of 2.5 x 109 CFU (colony forming unit)/ml and reaction volume of 500 µl, the efficiency of ultrasound DNA delivery (UDD) was 9.8 ± 2.3 x 10–6 transformants per cell, which was nine times more efficient than conjugation, and even four times greater than electroporation. We have also transferred pBBR1MCS2 into E. coli DH5 and P. fluorescens SBW25 with efficiencies of 1.16 ± 0.13 x 10–6 and 4.33 ± 0.78 x 10–6 transformants per cell, respectively. Low frequency UDD can be readily scaled up, allowing for the application of UDD not only in laboratory conditions but also on an industrial scale.

Item Type: Publication - Article
Digital Object Identifier (DOI): 10.1093/nar/gkm710
Programmes: CEH Programmes pre-2009 publications > Biodiversity
CEH Sections: Hails
ISSN: 0305-1048
Additional Information. Not used in RCUK Gateway to Research.: This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
NORA Subject Terms: Biology and Microbiology
Date made live: 18 Jan 2008 14:05
URI: http://nora.nerc.ac.uk/id/eprint/2117

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