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Group II introns as controllable gene targeting vectors for genetic manipulation of bacteria

Nature Biotechnology volume 19, pages 1162–1167 (2001)Cite this article

Abstract

Mobile group II introns can be retargeted to insert into virtually any desired DNA target. Here we show that retargeted group II introns can be used for highly specific chromosomal gene disruption in Escherichia coli and other bacteria at frequencies of 0.1–22%. Furthermore, the introns can be used to introduce targeted chromosomal breaks, which can be repaired by transformation with a homologous DNA fragment, enabling the introduction of point mutations. Because of their wide host range, mobile group II introns should be useful for genetic engineering and functional genomics in a wide variety of bacteria.

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Acknowledgements

This work was supported by NIH grant GM37949. We thank Stephanie Reeves, Shelley Payne and Rasika Harshey for Shigella and Salmonella strains and Marlene Belfort and Keith Derbyshire (Albany) for comments on the manuscript.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Institute for Cellular and Molecular Biology and Section of Molecular Genetics and Microbiology, School of Biological Sciences, University of Texas at Austin, Austin, 78712, TX

    Michael Karberg, Huatao Guo, Jin Zhong, Robert Coon, Jiri Perutka & Alan M. Lambowitz

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  1. Michael Karberg
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  2. Huatao Guo
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  3. Jin Zhong
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  4. Robert Coon
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  6. Alan M. Lambowitz
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Karberg, M., Guo, H., Zhong, J. et al. Group II introns as controllable gene targeting vectors for genetic manipulation of bacteria. Nat Biotechnol 19, 1162–1167 (2001). https://doi.org/10.1038/nbt1201-1162

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