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Application of In-Fusion™ Cloning for the Parallel Construction of E. coli Expression Vectors

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DNA Cloning and Assembly Methods

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1116))

Abstract

In-Fusion™ cloning is a flexible DNA ligase-independent cloning technology that has wide-ranging uses in molecular biology. In this chapter we describe the protocols used in the OPPF-UK to design and construct expression vectors using In-Fusion™. Our method for small scale expression screening in Escherichia coli of constructs generated by In-Fusion™ is also outlined.

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Acknowledgements

The Oxford Protein Production Facility-UK is supported by the UK Medical Research Council and the Biotechnology and Biology Research Council (MRC Grant MR/K018779/1). We thank Jo Nettleship for helpful discussions on E. coli secretion vectors.

Author information

Authors and Affiliations

  1. Oxford Protein Production Facility-UK, Oxfordshire, UK

    Louise E. Bird, Heather Rada, Jonathan M. Diprose & Raymond J. Owens

  2. Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Oxford, UK

    Louise E. Bird, Heather Rada, John Flanagan, Jonathan M. Diprose, Robert J. C. Gilbert & Raymond J. Owens

Authors
  1. Louise E. Bird
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  2. Heather Rada
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  3. John Flanagan
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  4. Jonathan M. Diprose
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  5. Robert J. C. Gilbert
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  6. Raymond J. Owens
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Editor information

Editors and Affiliations

  1. Norwegian University of Science and Technology, Trondheim, Norway

    Svein Valla

  2. Norwegian University of Science and Technology, Trondheim, Norway

    Rahmi Lale

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Bird, L.E., Rada, H., Flanagan, J., Diprose, J.M., Gilbert, R.J.C., Owens, R.J. (2014). Application of In-Fusion™ Cloning for the Parallel Construction of E. coli Expression Vectors. In: Valla, S., Lale, R. (eds) DNA Cloning and Assembly Methods. Methods in Molecular Biology, vol 1116. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-764-8_15

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  • DOI: https://doi.org/10.1007/978-1-62703-764-8_15

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-763-1

  • Online ISBN: 978-1-62703-764-8

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