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Stable Extracellular DNA: A Novel Substrate for Genetic Engineering that Mimics Horizontal Gene Transfer in Nature

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Extracellular Nucleic Acids

Part of the book series: Nucleic Acids and Molecular Biology ((NUCLEIC,volume 25))

Abstract

Molecular biology technology generally requires naked DNA purified in a test tube. However, extracellular DNA in the natural environment can contribute to breeding and growth as nutrition or genetic information. Transfer and maintenance of DNA is called horizontal gene transfer (HGT), a mechanism that has certainly played a role during evolution. The naked DNA received by particular microbes through natural transformation processes would almost certainly be extracellular. However, this form of transformation has been rarely studied compared with the other two major mechanisms of HGT: transduction and conjugation.

In this chapter, we present our recent discovery that plasmid DNA released from lysed Escherichia coli remains surprisingly stable in the environment. This extracellular DNA, of up to 100 kb and possibly larger, can undergo an HGT-like process into a recipient bacterium capable of natural transformation.

This form of DNA transfer, mimicking HGT, could facilitate the delivery of engineered genes without the need for biochemical purification, and permit experimental research into the mechanisms of HGT.

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Abbreviations

amp :

Ampicillin

BAC:

Bacterial artificial chromosome

bla :

β-Lactamase gene

bp:

Base pair(s)

cat :

Chloramphenicol acetyl transferase gene

ccc:

Covalently closed circular

cI:

cI Repressor gene

CMM:

Culture mix method

GFP:

Green fluorescent protein

GpBR:

Genomic pBR322 sequence

HGT:

Horizontal gene transfer

kb:

Kilobases

kbp:

Kilobase pairs

km :

Kanamycin resistance gene

tet :

Tetracycline resistance determinant gene

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Acknowledgments

We thank Dr. F. Maruyama of the Tokyo Institute of Technology for the helpful discussions.

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Authors and Affiliations

  1. Graduate School of Bioscience and Biotechnology, Department of Life Science, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama-shi, Kanagawa, 226-8501, Japan

    Shinya Kaneko

  2. Institute for Advanced Biosciences, Keio University, Nipponkoku, Tsuruoka-shi, Yamagata, 997-0017, Japan

    Mitsuhiro Itaya

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  1. Shinya Kaneko
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  2. Mitsuhiro Itaya
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Correspondence to Mitsuhiro Itaya .

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Editors and Affiliations

  1. , Dept. of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi-city, 441-8580, Japan

    Yo Kikuchi

  2. Fundamental Medicine, Institute of Chemical Biology and, Lavrentjeva Prospect 8, Novosibirsk, 630090, Russian Federation

    Elena Y. Rykova

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Kaneko, S., Itaya, M. (2010). Stable Extracellular DNA: A Novel Substrate for Genetic Engineering that Mimics Horizontal Gene Transfer in Nature. In: Kikuchi, Y., Rykova, E. (eds) Extracellular Nucleic Acids. Nucleic Acids and Molecular Biology, vol 25. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-12617-8_4

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