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Designer diatom episomes delivered by bacterial conjugation

Author

Listed:
  • Bogumil J. Karas

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute
    Present address: Designer Microbes Inc., London, Ontario, Canada.)

  • Rachel E. Diner

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute
    Scripps Institution of Oceanography, University of California San Diego)

  • Stephane C. Lefebvre

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute)

  • Jeff McQuaid

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute)

  • Alex P.R. Phillips

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute)

  • Chari M. Noddings

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute)

  • John K. Brunson

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute)

  • Ruben E. Valas

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute)

  • Thomas J. Deerinck

    (National Center for Microscopy and Imaging Research, University of California, San Diego)

  • Jelena Jablanovic

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute)

  • Jeroen T.F. Gillard

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute)

  • Karen Beeri

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute)

  • Mark H. Ellisman

    (National Center for Microscopy and Imaging Research, University of California, San Diego)

  • John I. Glass

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute)

  • Clyde A. Hutchison III

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute)

  • Hamilton O. Smith

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute)

  • J. Craig Venter

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute
    Microbial and Environmental Genomics Group, J. Craig Venter Institute)

  • Andrew E. Allen

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute
    Scripps Institution of Oceanography, University of California San Diego)

  • Christopher L. Dupont

    (Microbial and Environmental Genomics Group, J. Craig Venter Institute)

  • Philip D. Weyman

    (Synthetic Biology and Bioenergy Group, J. Craig Venter Institute)

Abstract

Eukaryotic microalgae hold great promise for the bioproduction of fuels and higher value chemicals. However, compared with model genetic organisms such as Escherichia coli and Saccharomyces cerevisiae, characterization of the complex biology and biochemistry of algae and strain improvement has been hampered by the inefficient genetic tools. To date, many algal species are transformable only via particle bombardment, and the introduced DNA is integrated randomly into the nuclear genome. Here we describe the first nuclear episomal vector for diatoms and a plasmid delivery method via conjugation from Escherichia coli to the diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. We identify a yeast-derived sequence that enables stable episome replication in these diatoms even in the absence of antibiotic selection and show that episomes are maintained as closed circles at copy number equivalent to native chromosomes. This highly efficient genetic system facilitates high-throughput functional characterization of algal genes and accelerates molecular phytoplankton research.

Suggested Citation

  • Bogumil J. Karas & Rachel E. Diner & Stephane C. Lefebvre & Jeff McQuaid & Alex P.R. Phillips & Chari M. Noddings & John K. Brunson & Ruben E. Valas & Thomas J. Deerinck & Jelena Jablanovic & Jeroen T, 2015. "Designer diatom episomes delivered by bacterial conjugation," Nature Communications, Nature, vol. 6(1), pages 1-10, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms7925
    DOI: 10.1038/ncomms7925
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    Cited by:

    1. Huan Zhang & Xiaofeng Xiong & Kangning Guo & Mengyuan Zheng & Tianjun Cao & Yuqing Yang & Jiaojiao Song & Jie Cen & Jiahuan Zhang & Yanyou Jiang & Shan Feng & Lijin Tian & Xiaobo Li, 2024. "A rapid aureochrome opto-switch enables diatom acclimation to dynamic light," Nature Communications, Nature, vol. 15(1), pages 1-18, December.
    2. E. J. L. Walker & M. Pampuch & L. Deng & Y. Li & G. Tran & T. Mock & B. J. Karas, 2026. "Breaking the cell wall for efficient DNA delivery to diatoms," Nature Communications, Nature, vol. 17(1), pages 1-18, December.

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