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Genetic Engineering of Cyanobacteria: Design, Implementation, and Characterization of Recombinant Synechocystis sp. PCC 6803

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Microbial Metabolic Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1927))

Abstract

Synechocystis sp. PCC 6803 is a model cyanobacterium which has been investigated to produce a variety of fuels and chemicals. Genetic mutations are of interest for studying photosynthesis and engineering chemical production. Here, methods for culturing, preserving, and genetically transforming Synechocystis sp. PCC 6803 are detailed including methods to test promoter strength using the green fluorescent protein reporter. Furthermore, a method for markerless transformation of chromosomal DNA is presented. Sufficient details are provided to enable application by the novice investigator.

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Acknowledgments

The authors acknowledge funding for this work by the National Science Foundation (NSF awards 1336236 and 1332404). Stevan Albers and Yi Ern Cheah contributed to this work by developing the protocols for transforming Synechocystis 6803 used in our lab and teaching us these methods.

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Author notes

  1. Jacob Sebesta and Allison Werner contributed equally to this chapter.

Authors and Affiliations

  1. Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, CO, USA

    Jacob Sebesta & Christie Ann Marie Peebles

  2. Cell and Molecular Biology Graduate Program, Colorado State University, Fort Collins, CO, USA

    Allison Werner & Christie Ann Marie Peebles

Authors
  1. Jacob Sebesta
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  2. Allison Werner
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  3. Christie Ann Marie Peebles
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Corresponding author

Correspondence to Christie Ann Marie Peebles .

Editor information

Editors and Affiliations

  1. Manus Bio, Cambridge, MA, USA

    Christine Nicole S. Santos

  2. Manus Bio, Cambridge, MA, USA

    Parayil Kumaran Ajikumar

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Sebesta, J., Werner, A., Peebles, C.A.M. (2019). Genetic Engineering of Cyanobacteria: Design, Implementation, and Characterization of Recombinant Synechocystis sp. PCC 6803. In: Santos, C., Ajikumar, P. (eds) Microbial Metabolic Engineering. Methods in Molecular Biology, vol 1927. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9142-6_10

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  • DOI: https://doi.org/10.1007/978-1-4939-9142-6_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9141-9

  • Online ISBN: 978-1-4939-9142-6

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