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Toward a genetic system in the marine cyanobacterium Prochlorococcus

View ORCID ProfileRaphaël Laurenceau, Christina Bliem, Marcia S. Osburne, Jamie W. Becker, Steven J. Biller, Andres Cubillos-Ruiz, Sallie W. Chisholm
doi: https://doi.org/10.1101/820027
Raphaël Laurenceau
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA
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  • ORCID record for Raphaël Laurenceau
  • For correspondence: raphaellaurenceau@gmail.com chisholm@mit.edu
Christina Bliem
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA
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Marcia S. Osburne
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, USADepartment of Molecular Biology and Microbiology Tufts University School of Medicine, Boston, MA, USA
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Jamie W. Becker
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, USADepartment of Biology, Haverford College, Haverford, PA, USA
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Steven J. Biller
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, USADepartment of Biological Sciences, Wellesley College, Wellesley, MA, USA
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Andres Cubillos-Ruiz
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, USAInstitute for Medical Engineering and Science, Department of Biological Engineering, and Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, USABroad Institute of MIT and Harvard, Cambridge, MA, USAWyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA
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Sallie W. Chisholm
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, USADepartment of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA
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  • For correspondence: raphaellaurenceau@gmail.com chisholm@mit.edu
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ABSTRACT

As the smallest and most abundant primary producer in the oceans, the cyanobacterium Prochlorococcus is of interest to diverse branches of science. For the past 30 years, research on this minimal phototroph has led to a growing understanding of biological organization across multiple scales, from the genome to the global ocean ecosystem. Progress in understanding drivers of its diversity and ecology, as well as molecular mechanisms underpinning its streamlined simplicity, has been hampered by the inability to manipulate these cells genetically.

Multiple attempts have been made to develop an efficient genetic transformation method for Prochlorococcus over the years; all have been unsuccessful to date, despite some success with their close relative, Synechococcus. To avoid the pursuit of unproductive paths, we report here what has not worked in our hands, as well as our progress developing a method to screen the most efficient electroporation parameters for optimal DNA delivery into Prochlorococcus cells. We also report a novel protocol for obtaining axenic colonies and a new method for differentiating live and dead cells. The electroporation method can be used to optimize DNA delivery into any bacterium, making it a useful tool for advancing transformation systems in other genetically recalcitrant microorganisms.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted October 25, 2019.
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Toward a genetic system in the marine cyanobacterium Prochlorococcus
Raphaël Laurenceau, Christina Bliem, Marcia S. Osburne, Jamie W. Becker, Steven J. Biller, Andres Cubillos-Ruiz, Sallie W. Chisholm
bioRxiv 820027; doi: https://doi.org/10.1101/820027
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Toward a genetic system in the marine cyanobacterium Prochlorococcus
Raphaël Laurenceau, Christina Bliem, Marcia S. Osburne, Jamie W. Becker, Steven J. Biller, Andres Cubillos-Ruiz, Sallie W. Chisholm
bioRxiv 820027; doi: https://doi.org/10.1101/820027

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