High-Efficiency Transformation of Chlamydomonas reinhardtii by Electroporation | Zendy

Kosuke Shimogawara | Zendy; Shoko Fujiwara | Zendy; Arthur Grossman | Zendy; Hideaki Usuda | Zendy

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High-Efficiency Transformation of Chlamydomonas reinhardtii by Electroporation

Author(s) -

Kosuke Shimogawara,

Shoko Fujiwara,

Arthur Grossman,

Hideaki Usuda

Publication year - 1998

Publication title -

genetics

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 2.792

H-Index - 246

eISSN - 1943-2631

pISSN - 0016-6731

DOI - 10.1093/genetics/148.4.1821

Subject(s) - chlamydomonas reinhardtii , electroporation , chlamydomonas , biology , transformation (genetics) , complementation , plasmid , mutant , transformation efficiency , dna , exogenous dna , genetics , microbiology and biotechnology , gene , agrobacterium

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 × 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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