Genetic Transformation

  • Peter G. Kroth
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 390)

Diatoms are unicellular photoautotrophic eukaryotes that play an important role in ecology by fixing large amounts of CO2 in the oceans. Because they evolved by secondary endocytobiosis—a process of uptake of a eukaryotic alga into another eukaryotic cell—they have a rather unusual cell biology and genetic constitution. Because the preparation of organelles is rather difficult as a result of the cytosolic structures, genetic transformation and expression of preproteins fused to green fluorescent protein (GFP) became one of the major tools to analyze subcellular localization of proteins in diatoms. Meanwhile several groups successfully attempted to develop genetic transformation protocols for diatoms. These methods are based on ‘‘biolistic’’ DNA delivery via a particle gun and allow the introduction and expression of foreign genes in the algae. Here a protocol for the genetic transformation of the diatom Phaeodactylum tricornutum is described as well as the subsequent characterization of the transformants.

Key Words

Diatom transformation screening targeting microscopy. 
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Notes

Acknowledgements

The author would like to thank Ansgar Gruber and Doris Ballert for careful reading of the manuscript and helpful suggestions. This work was supported by the German Research Foundation (DFG, project Kr 1661/3-1), the European Community (MARGENES, contract QLRT-2001-01226), and the University of Konstanz.

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Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Peter G. Kroth
    • 1
  1. 1.Department of BiologyUniversity of KonstanzKonstanzGermany

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