HIGH-EFFICIENCY TRANSFORMATION OF CHLAMYDOMONAS-REINHARDTII BY ELECTROPORATION

We have established a high-efficiency method for transforming the unic ellular, green alga Chlamydomonas reinhardtii by electroporation. Elec troporation 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 fo r the introduction of exogenous DNA. The conditions that were varied i ncluded osmolarity, temperature, concentration of exogenous DNA, volta ge and capacitance. Following optimization, the maximum transformation frequency obtained was 2 x 10(5) transformants per mu g of DNA; this frequency is two orders of magnitude higher than obtained with the cur rent standard method using glass beads to introduce exogenous DNA. The electroporation procedure described in this article is of general uti lity, and makes it feasible to isolate genes by direct complementation of Chlamydomonas reinhardtii mutants.