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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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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Short Communication

Polyethylene Glycol (PEG)-Mediated Transient Gene Expression in a Red Alga, Cyanidioschyzon merolae 10D

Mio Ohnuma1, Takashi Yokoyama2, Takayuki Inouye2, Yasuhiko Sekine2 and Kan Tanaka1,*

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

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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 Proc. Natl. Acad. Sci. USAHome page
S. Imamura, Y. Kanesaki, M. Ohnuma, T. Inouye, Y. Sekine, T. Fujiwara, T. Kuroiwa, and K. Tanaka
R2R3-type MYB transcription factor, CmMYB1, is a central nitrogen assimilation regulator in Cyanidioschyzon merolae
PNAS, July 28, 2009; 106(30): 12548 - 12553.
[Abstract] [Full Text] [PDF]


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