Abstract
The unicellular red alga Cyanidioschyzon merolae shows simple cell structure, containing one nucleus, one mitochondrion and one plastid. In order to use this organism as a model cell, we are developing transformation techniques to facilitate functional genomics.
Here, we show that DNA is easily introduced into C. merolae cells by a polyethylene glycol (PEG)-mediated protocol. β-tubulin gene of C. merolae was cloned on a plasmid and a hemagglutinin (HA) tag was added at the C-terminus. This plasmid was introduced into C. merolae cells by a PEG-mediated protocol. At 24 h after transformation, intracellular localization of the tagged protein was detected by anti-HA immunocytochemistry. By considering the PEG molecular weights and the final concentration, 30% PEG4000 was found as the optimum condition. The number of apparently fluorescent cells was counted, and the transformation frequency was calculated as 2-10% based on the number of DAPI-stained cells.
