Plant and Cell Physiology Advance Access originally published online on November 14, 2007
Plant and Cell Physiology 2008 49(1):117-120; doi:10.1093/pcp/pcm157
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Short Communication |
Polyethylene Glycol (PEG)-Mediated Transient Gene Expression in a Red Alga, Cyanidioschyzon merolae 10D
1Institute of Molecular and Cellular Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-0032 Japan
2Department of Life Science, College of Science, Rikkyo (St Paul's) University, Nishiikebukuro, Tokyo, 171-8501 Japan
*Corresponding author: E-mail, kntanaka{at}iam.u-tokyo.ac.jp; Fax, +81-3-5841-8476.
| Abstract |
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DNA introduction into cells is an essential technique for molecular genetic analysis. Here, we show that DNA is easily introduced into cells of the unicellular red alga Cyanidioschyzon merolae by a polyethylene glycol (PEG)-mediated protocol. In this study, the β-tubulin gene of C. merolae was cloned on a plasmid and a hemagglutinin (HA) tag then added at the C-terminus. This plasmid was then introduced into C. merolae cells by a PEG-mediated transformation protocol. At 24 h after PEG-mediated transformation, intracellular localization of the tagged protein was detected by anti-HA immunocytochemistry, indicating the utility of this transient expression system for molecular genetic analyses.
Keywords: Cyanidioschyzon merolae - Hemagglutinin - Polyethylene glycol - Transformation - Transient expression system
Abbreviations: DAPI, 4', 6-diamidino-2-phenylindole; 5-FOA, 5-fluoroorotic acid; HA, hemagglutinin; MA medium, modified Allen's medium; PEG, polyethylene glycol.
(Received September 21, 2007; Accepted November 8, 2007)
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