Designer diatom episomes delivered by bacterial conjugation

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.F. Gillard, Karen Beeri, Mark H. Ellisman, John I. Glass, Clyde A. Hutchison, Hamilton O. Smith, J. Craig Venter, Andrew E. Allen, Christopher L. Dupont and Philip D. Weyman ()
Additional contact information
Bogumil J. Karas: Synthetic Biology and Bioenergy Group, J. Craig Venter Institute
Rachel E. Diner: Microbial and Environmental Genomics Group, J. Craig Venter Institute
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: 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
Andrew E. Allen: Microbial and Environmental Genomics Group, J. Craig Venter Institute
Christopher L. Dupont: Microbial and Environmental Genomics Group, J. Craig Venter Institute
Philip D. Weyman: Synthetic Biology and Bioenergy Group, J. Craig Venter Institute

Nature Communications, 2015, vol. 6, issue 1, 1-10

Abstract: 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.

Date: 2015
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DOI: 10.1038/ncomms7925

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