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1 Department of Zoology, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan
2 Institute of Chemical Research, Kyoto University, Uji 611-0011, Japan
3 Department of Biological Sciences, Tokyo Institute of Technology, Yokohama 226-8501, Japan
4 Department of Biology, Faculty of Science, Konan University, Kobe 658-8501, Japan
5 High Technology Research Center, Konan University, Kobe 658-8501, Japan
*Author for correspondence (e-mail: kwmakabe{at}ascidian.zool.kyoto-u.ac.jp)
Accepted April 16, 2001
The ascidian egg is a well-known mosaic egg. In order to investigate the molecular nature of the maternal genetic information stored in the egg, we have prepared cDNAs from the mRNAs in the fertilized eggs of the ascidian, Halocynthia roretzi. The cDNAs of the ascidian embryo were sequenced, and the localization of individual mRNA was examined in staged embryos by whole-mount in situ hybridization. The data obtained were stored in the database MAGEST (http://www.genome.ad.jp/magest) and further analyzed. A total of 4240 cDNA clones were found to represent 2221 gene transcripts, including at least 934 different protein-coding sequences. The mRNA population of the egg consisted of a low prevalence, high complexity sequence set. The majority of the clones were of the rare sequence class, and of these, 42% of the clones showed significant matches with known peptides, mainly consisting of proteins with housekeeping functions such as metabolism and cell division. In addition, we found cDNAs encoding components involved in different signal transduction pathways and cDNAs encoding nucleotide-binding proteins. Large-scale analyses of the distribution of the RNA corresponding to each cDNA in the eight-cell, 110-cell and early tailbud embryos were simultaneously carried out. These analyses revealed that a small fraction of the maternal RNAs were localized in the eight-cell embryo, and that 7.9% of the clones were exclusively maternal, while 40.6% of the maternal clones showed expression in the later stages. This study provides global insights about the genes expressed during early development.
Key words: Ascidian, Maternal RNA, cDNA analysis, EST, Expression profile, Database
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