Gene Expression Profiling of Coccolith-Bearing Cells and Naked Cells in Haptophyte Pleurochrysis haptonemofera with a cDNA Macroarray System
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- 作者：Shoko Fujiwara ; Yasutaka Hirokawa ; Yukiko Takatsuka ; Kunihiro Suda ; Erika Asamizu ; Takatoshi Takayanagi ; Daisuke Shibata ; Satoshi Tabata and Mikio Tsuzuki
- 关键词：calcification ; carbonic anhydrase ; cDNA macroarray ; coccolithophorid ; Pleurochrysis haptonemofera
- 刊名：Marine Biotechnology
- 出版年：2007
- 出版时间：October, 2007
- 年：2007
- 卷：9
- 期：5
- 页码：550-560
- 全文大小：380 KB
- 刊物类别：Earth and Environmental Science
- 刊物主题：Earth sciences
Oceanography - 出版者：Springer New York
- ISSN：1436-2236
文摘
Pleurochrysis haptonemofera is a unicellular marine coccolithophorid that has calcified scales, coccoliths, on the cell surface. Some coccolithophorids including P. haptonemofera have a coccolith-bearing stage and a naked stage in their life cycles. To characterize genes involved in the coccolithogenesis, we generated a total of 9550 expressed sequence tags (EST) from a normalized cDNA library that was prepared using both coccolith-bearing cells (C-cells) and naked cells (N-cells), constructed a cDNA macroarray using the EST clones, and then analyzed the gene expression specificity in C-cells and N-cells. When cDNA clones whose expression ratio exceeded 3-fold were selected, as many as 180 clones were identified as C-cell-specific ones, while only 12 were found to be N-cell-specific ones. These clones were sequenced, assembled, and homology-searched against a public nonredundant protein database. As a result, they were grouped into 54 C-cell-specific and 6 N-cell-specific genes, and 59% and 50% of these genes exhibited significant similarity to those of other known proteins, respectively. To assess mRNA expression further, Northern hybridization was performed for 12 of the C-cell-specific genes and one of the N-cell-specific ones. These clones, together with the new cDNA macroarray, will provide a powerful tool for the future genome-wide functional analysis of uncharacterized genes related to the regulation of the calcification and life cycle of coccolithophorids.