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Fluorescent Differential Display Identifies Circadian Clock-Regulated Genes in Arabidopsis thaliana

Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037, USA; Novartis Agricultural Discovery Institute, Inc., 15 Merryfield Row, Suite 100, San Diego, CA92121, USA

Takamichi Muramatsu

Hitachi Advanced Research Laboratory, Hatoyama, Saitama 350 0395, Japan; Hitachi Life Science Group, Hatoyama, Saitama 350-0395, Japan

Masaki Furuya

Hitachi Advanced Research Laboratory, Hatoyama, Saitama 350 0395, Japan

Steve A. Kay

Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037, USA

Circadian rhythms in gene expression were first observed in plants more than 13 years ago, but the underlying mechanism controlling rhythmic gene expression is still not understood. The isolation of novel circadian clock-controlled genes (ccgs) is likely to provide new tools for studying circadian rhythms. Fluorescent differential display (FDD) was used to screen Arabidopsis thaliana mRNAs for cycling transcripts. Seventy PCR primer pairs were screened, and 17 different cycling bands were observed out of an estimated 10,500 bands screened. The identities of 10 bands were determined, and the rhythmic gene expression was confirmed using northern blot analysis. The 10 cycling bands represent 7 different genes, 6 of which are present in the databases and 1 that does not match anything in current databases. The rhythmic expression of the 7 genes is composed of four distinct phases of clock regulation. The results demonstrate that FDD can be used to isolate ccgs. The genes identified in this screen range from known A. thaliana ccgs, as well as genes shown to be clock controlled in other plant species, to a novel gene that may encode a pioneer protein. Further study of these ccgs is likely to increase our understanding of circadian-regulated gene expression.

Key Words: circadian clock-controlled gene expression • fluorescent differential display • Arabidopsis thaliana

Journal of Biological Rhythms, Vol. 15, No. 3, 208-217 (2000)
DOI: 10.1177/074873040001500302


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