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Bacteriophage φC31 Integrase Mediated Transgenesis in Xenopus laevis for Protein Expression at Endogenous Levels

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Book cover Microinjection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 518))

Abstract

Bacteriophage φC31 inserts its genome into that of its host bacterium via the integrase enzyme which catalyzes recombination between a phage attachment site (attP) and a bacterial attachment site (attB). Integrase requires no accessory factors, has a high efficiency of recombination, and does not need perfect sequence fidelity for recognition and recombination between these attachment sites. These imperfect attachment sites, or pseudo-attachment sites, are present in many organisms and have been used to insert transgenes in a variety of species. Here we describe the φC31 integrase approach to make transgenic Xenopus laevis embryos.

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Acknowledgements

We would like to thank Professor Michele Calos for providing the pET11phiC31poly(A) plasmid, Professor Gary Felsenfeld for providing the HS4 insulator sequences, and Paul Kreig for providing the gamma crystallin lens promoter. This work was supported by funding from the NIH (GM069944 and DC007481). Bryan Allen is a student in the Medical Scientist Training Program at the Roy J. and Lucille A Carver College of Medicine, University of Iowa.

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Authors and Affiliations

  1. Department of Biochemistry, University of Iowa, Iowa City, IA, USA

    Bryan G. Allen & Daniel L. Weeks

Authors
  1. Bryan G. Allen
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  2. Daniel L. Weeks
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Editors and Affiliations

  1. Dept. Biological Sciences, Florida Institute of Technology, W. University Boulevard 150, Melbourne, 32901, U.S.A.

    David J. Carroll

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Allen, B.G., Weeks, D.L. (2009). Bacteriophage φC31 Integrase Mediated Transgenesis in Xenopus laevis for Protein Expression at Endogenous Levels. In: Carroll, D. (eds) Microinjection. Methods in Molecular Biology, vol 518. Humana Press. https://doi.org/10.1007/978-1-59745-202-1_9

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  • DOI: https://doi.org/10.1007/978-1-59745-202-1_9

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-884-3

  • Online ISBN: 978-1-59745-202-1

  • eBook Packages: Springer Protocols

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