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Genetics, Vol. 148, 1821-1828, April 1998, Copyright © 1998

High-Efficiency Transformation of Chlamydomonas reinhardtii by Electroporation

Kosuke Shimogawaraa, Shoko Fujiwarab, Arthur Grossmanc, and Hideaki Usudaa
a Laboratory of Chemistry, Teikyo University School of Medicine, Hachioji, Tokyo, 192-03 Japan,
b School of Life Science, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo, 192-03 Japan
c Department of Plant Biology, Carnegie Institution of Washington, Stanford, CA 94305

Corresponding author: Kosuke Shimogawara, Laboratory of Chemistry, Teikyo University School of Medicine, 359 Ohtsuka, Hachioji, Tokyo 192-0395, Japan, kosuke{at}main.teikyo-u.ac.jp (E-mail).

Communicating editor: J. J. LOROS

We have established a high-efficiency method for transforming the unicellular, green alga Chlamydomonas reinhardtii by electroporation. Electroporation of strains CC3395 and CC425, cell wall-less mutants devoid of argininosuccinate lyase (encoded by ARG7 ), in the presence of the plasmid pJD67 (which contains ARG7 ) was used to optimize conditions for the introduction of exogenous DNA. The conditions that were varied included osmolarity, temperature, concentration of exogenous DNA, voltage and capacitance. Following optimization, the maximum transformation frequency obtained was 2 x 105 transformants per µg of DNA; this frequency is two orders of magnitude higher than obtained with the current standard method using glass beads to introduce exogenous DNA. The electroporation procedure described in this article is of general utility, and makes it feasible to isolate genes by direct complementation of Chlamydomonas reinhardtii mutants.




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